For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/assay-kits/complex-ii-enzyme-activity-microplate-assay-kit-ab109908.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Metabolism Mitochondrial
Share by email

Complex II Enzyme Activity Microplate Assay Kit (ab109908)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (51)References (96)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

- Complex II Enzyme Activity Microplate Assay Kit (ab109908)
  • - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
  • Functional Studies - Complex II Enzyme Activity Microplate Assay Kit (ab109908)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 3 hr
  • Sample type: Cell Lysate, Purified mitochondria, Tissue Lysate

You may also be interested in

Assay
Product image
Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909)
Lysate
Bovine Heart Mitochondria (ab110338)
Biochemical
Product image
Atpenin A5, ubiquinone-binding site mitochondrial complex II inhibitor (ab144194)

View more associated products

Overview

  • Product name

    Complex II Enzyme Activity Microplate Assay Kit
    See all Complex II kits
  • Detection method

    Colorimetric
  • Sample type

    Cell Lysate, Tissue Lysate, Purified mitochondria
  • Assay type

    Enzyme activity
  • Assay time

    3h 00m
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    Complex II Enzyme Activity Microplate Assay Kit is designed for determining the Complex II activity in a human, mouse, rat or bovine sample. Each of the 96 wells in the kit has been coated with an anti-Complex II monoclonal antibody (mAb) which purifies the enzyme from a complex sample such as mitochondria, tissue homogenate or cell lysate. After this in-well purification the production of ubiquinol by the enzyme is coupled to the reduction of the dye DCPIP (2,6-diclorophenolindophenol) and a decreases in its absorbance at 600 nm, which in turn recycles the substrate ubiquinone.

  • Notes

    Succinate, Ubiquinone 2, DCPIP and Lipid?Phospholipd Mix should be stored at -80°C. All other components should be stored at 4°C.

    Related products

    Review the mitochondrial assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 96 tests
    10X Blocking Solution 1 x 5ml
    Detergent 2 x 1ml
    20X Buffer 1 x 15ml
    Complex II Activity Buffer 1 x 25ml
    DCPIP/DCIP 1 x 250µl
    Phospholipids 1 x 6ml
    Pre-coated 96-well microplate (12 strips) 1 unit
    Succinate 1 x 500µl
    Ubiquinone 2 1 x 60µl
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Other Metabolism Assay
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
  • Relevance

    Complex II is also called succinate ubiquinone oxidoreductase or more commonly succinate dehydrogenase complex. This complex is composed of four nuclear encoded subunits and contains a flavin (FAD), non-heme iron centers and a b-type cytochrome as prosthetic groups. It is both a component of the electron transport chain and an enzyme of the Krebs cycle. Complex II deficiencies are seen in OXPHOS genetic disease and found in a type of cancer called paraganglioma.
  • Alternative names

    • SDH
    • Succinate coenzyme Q reductase
    • Succinate dehydrogenase
  • Database links

    • Entrez Gene: 6390 Human
    • Entrez Gene: 6389 Human
    • Entrez Gene: 6391 Human
    • SwissProt: P31040 Human
    • SwissProt: P21912 Human
    • SwissProt: Q99643 Human
    • SwissProt: O14521 Human
    • SwissProt: 6392 Human

    Associated products

    • Positive Controls

      • Bovine Heart Mitochondria (ab110338)
      • Rat liver tissue lysate - mitochondrial extract (ab110346)
      • Rat heart tissue lysate - mitochondrial extract (ab110347)
      • Rat brain tissue lysate - mitochondrial extract (ab110348)
      • Mouse liver tissue lysate - mitochondrial extract (ab110349)
      • Mouse heart tissue lysate - mitochondrial extract (ab110350)
      • Mouse brain tissue lysate - mitochondrial extract (ab110351)

    Images

    • - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      Figure 2. This assay is compatible with different sample types such as mitochondria, tissue or cell lysates and in multiple species including human and rodent samples. Typical linear range data are shown for ab109908.
    • - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      Figure 1. Example of raw data. Note the lag period before activity. Also note the activity of mitochondria (BHM, bovine heart mitochondria) is higher than whole cell lysate (HepG2, human hepatoblastoma) and the reaction ends at >1600 seconds because the substrates are used up.
    • Functional Studies - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      Functional Studies - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
      Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.

      Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, substrate is added, and enzyme activity is analyzed by measuring the change in absorbance of either the substrate or the product of the reaction (depending upon which enzyme is being analyzed). By analyzing the enzyme's activity in an isolated context, outside of the cell and free from any other variables, an accurate measurement of the enzyme's functional state can be understood.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (96)

    Publishing research using ab109908? Please let us know so that we can cite the reference in this datasheet.

    ab109908 has been referenced in 96 publications.

    • Rajab BS  et al. Differential remodelling of mitochondrial subpopulations and mitochondrial dysfunction are a feature of early stage diabetes. Sci Rep 12:978 (2022). PubMed: 35046471
    • Han G  et al. Dihuang-Yinzi Alleviates Cognition Deficits via Targeting Energy-Related Metabolism in an Alzheimer Mouse Model as Demonstrated by Integration of Metabolomics and Network Pharmacology. Front Aging Neurosci 14:873929 (2022). PubMed: 35431901
    • Erdem A  et al. Inhibition of the succinyl dehydrogenase complex in acute myeloid leukemia leads to a lactate-fuelled respiratory metabolic vulnerability. Nat Commun 13:2013 (2022). PubMed: 35440568
    • Dong XQ  et al. Implication of a novel truncating mutation in titin as a cause of autosomal dominant left ventricular noncompaction. J Geriatr Cardiol 19:301-314 (2022). PubMed: 35572216
    • Zou R  et al. Empagliflozin attenuates cardiac microvascular ischemia/reperfusion injury through improving mitochondrial homeostasis. Cardiovasc Diabetol 21:106 (2022). PubMed: 35705980
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-53 of 53 Abreviews or Q&A

    control of mitochondrial function and cell growth by the atypical cadherin Fat1

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    Easy to follow instruction and standardize using cell lysate. Good sensitivity with microplate reader
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 27 2018

    Short-term cigarette smoke exposure induces reversible changes in energy metabolism and cellular redox status independent of inflammatory responses in mouse lungs.

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    30ug Lung mitochondrion samples from mice in buffer containing 250 mM sucrose, 20 mM HEPES, 1 mM EDTA, 1 mM EGTA, 0.25% protease inhibitor, and 0.5% BSA, pH 7.4 were used for the assay.
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Amit Agarwal

    Verified customer

    Submitted Jul 24 2013

    Question

    On page 8 steps 3 and 4, the protein concentration needs to be determined before the detergent is added. How do you ensure the samples are at 5.5 mg/mL?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 15 2012

    Answer

    Thank you very much for your call today and for your patience while I have been in touch with the lab scientists regarding this enquiry.

    If you're using a tissue homogenate or whole mitochondria sample, before extraction with the kit’s detergent (lauryl maltoside)the protein concentration of the sampleshould be 5.5mg/mL.A way of doing this is as follows:

    Sacrifice a small portion of the sample (10 – 20uL), solublize it with any ionic detergent of choice and then determine protein concentration in this small aliquot. That number will be representative (taking into account any potential dilutions made) of the protein concentration of the original sample.

    If the equivalent concentration was higher than 5.5mg/mL add more buffer accordingly. If the equivalent concentration was lower than 5.5mg/mL centrifuge the whole tissue homogenate or whole mitochondria at high speed. This will generate a pellet which can then be resuspended in a smaller volume.

    Once the sample is at the right concentration,you can add the kit’s detergent and follow the instructions



    We have a booklet that explains in more detail sample extraction. This booklet can be found in the following link:

    http://www.mitosciences.com/PDF/mitosciences-sample-preparation-guide.pdf

    I hope that this information will be useful, but if please let me know if you have any further questions or need anything else and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Aug 15 2012

    Question

    I'm using this kit, but I haven't determined the protein concentration. I'm trying to find the right cell number to use. I'm using 100,000, 1 million, and 10 million lymphoma cells. I tried something similar before, but the assay didn't work (no signal above negative control). Do I have to dilute my sample in the incubation buffer in order for the assay to work? I want to make sure it's concentrated enough, and I haven't measured it to the recommended 1.2 mg/ml, so is there some sort of ratio I could follow (i.e. 50 ul sample, 10 ul incubation buffer)?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 31 2012

    Answer

    Thank you for your inquiry.


    The assays can be optimized by cell number, but since all cells are different sizes we use total protein mass.


    In general large cells such as fibroblast, HeLa etc are about 0.25 mg/million. So for a normal cell, 10 million would work.

    However lymphoblasts are almost always very small cells and would require even more cells.

    One solution is to resuspend the cell pellet to 10 volumes with extraction buffer to get a protein concentration of 2-5 mg/mL.

    So to get a 200 uL sample you need to start with 20 uL cell pellet + 180 uL lysis buffer.

    We would always recommend using a positive control – a human cell line e.g. HeLa or one of our mitochondrial samples (below)

    Name
    Species
    Applications
    Amounts
    MitoSciences code
    Abcam code

    https://www.abcam.com/ab110351
    Ms
    Immunocapture, BNPage
    2 mg
    MS822
    https://www.abcam.com/ab110351

    https://www.abcam.com/ab110348
    Rat
    Immunocapture, BNPage
    2 mg
    MS819
    https://www.abcam.com/ab110348

    https://www.abcam.com/ab110338
    Cow
    Immunocapture, BNPage
    2 mg
    MS802
    https://www.abcam.com/ab110338

    https://www.abcam.com/ab110350
    Ms
    Immunocapture, BNPage
    2 mg
    MS821
    https://www.abcam.com/ab110350

    https://www.abcam.com/ab110347
    Rat
    Immunocapture, BNPage
    2 mg
    MS818
    https://www.abcam.com/ab110347

    https://www.abcam.com/ab110349
    Ms
    Immunocapture, BNPage
    2 mg
    MS820
    https://www.abcam.com/ab110349

    https://www.abcam.com/ab110346
    Rat
    Immunocapture, BNPage
    2 mg
    MS811
    https://www.abcam.com/ab110346


    I hope this information helps. Please contact us with any other questions.

    Read More

    Abcam Scientific Support

    Answered on Jan 31 2012

    Question

    I am planning to investigate the OXPHOS pathway in rat heart tissue. I have looked at your complex assays (ab109721, ab109908, ab109911 and ab109716) and your mitochondrial isolation kit for tissue (ab110168) and had a few questions: 1. Are the mitochondria isolated from the kit suitable for use on all of the above assays? 2. Is there any further treatment that is required before the isolated mitochondria can be used in each assay (i.e. incubation with detergent to isolate protein complex) or are they used directly into the plate? Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2015

    Answer


    1) Yes, the mitochondrial isoaltion kit is suitable for use with all kits listed.

    2) Suspend the prepared mitochondria to 5.5 mg/mL as described in the attached document and follow from step 5 (adding detergent to mitochondia) to generate an extract suitable for all kits.

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Nov 16 2015

    Question

    Hi,
     
    With regards to using Malonate (Buffered Malonic Acid) for an inhibitor of Complex II what concentration and pH is recommended? Thanks.
     

    Read More

    Abcam community

    Verified customer

    Asked on Apr 14 2014

    Answer

    According to the literature, malonate inhibits complex II with an IC50 of ˜40 μM (PMCID: PMC2507763). Please let me know if you have any further questions!

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Apr 14 2014

    Question



    Inquiry: Hello, I recently purchased this kit, and I have a few questions before proceeding with my pilot experiment. 1.) Should I keep samples (cells) on ice while I am resuspending them? 2.) Should I add a protease inhibitor to the detergent? 3.) I have isolated mito for a positive control (to ensure the kit works). However, I am trying to come up with a positive control for increased activity of Complex II. Do you know of any compounds or even amino acid mutations that allow for increased activity of Complex II? 4.) I would like to use a negative control- that decreases activity of Complex II. In the protocol, you mention TTFA as an inhibitor of Complex II. Can I add the TTFA to the activity solution and decrease the amount of activity buffer used? 5.) Do you know if post-translational modifications like acetylation are maintained on Complex II bound to the mAb? Thanks for all your help!

    Read More

    Abcam community

    Verified customer

    Asked on Feb 17 2014

    Answer

    Thank you for contacting us.

    Yes, you should keep samples on ice, and also add a protease inhibitor to the detergent.

    Yes TTFA can be added and should decrease the activity of Complex II.
    However TTFA is not as specific/effective as some of the other OXPHOS inhibitors. Malonate (Buffered malonic acid) could also be used.
    Acetylation should be preserved. The capture antibody is a mouse antibody and we have been able to see acetylation by adding a rabbit anti-acetyl lysine antibody ab21623 and Goat anti-rabbit HRP to the plate.
    https://www.abcam.com/acetyl-Lysine-antibody-ChIP-Grade-ab21623.html


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    -----------------------------------------------------------------------------------------------

    Do you wish running an ELISA was fast and easy? Check out our new SimpleStep™ ELISA, a single wash, colorimetric sandwich ELISA assay.

    Discover more at www.abcam.com/simplestep

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Feb 17 2014

    Question

    Phone call: How to analyse the concentration of cell lysates?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 20 2013

    Answer

    Pipette up and down cells several time for complete lysis and then determine the protein concentration using BCA or Bradford method. Once protein concentration is known then please follow rest of the procedure.

    20 million of average sized cells per ml will give 5mg/ml of protein.

    Read More

    Padamjeet Singh

    Abcam Scientific Support

    Answered on Nov 20 2013

    Question

    Customer is trying to calculate rate of activity in mM/min/mg protein as seen in fig.11 of  paper Li C  et al. Free Radic Biol Med 60C:29-40 (2013). (PubMed: 23376468, http://www.sciencedirect.com/science/article/pii/S0891584913000233) In order to do this they are asking if we can provide the concnetration of DCIP provided in this kit.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 12 2013

    Answer

    The DCIP concentration is 2.5mM.

    Read More

    Keith B

    Abcam Scientific Support

    Answered on Jul 12 2013

    Question

    Do you have any recommendations for OXPHOS assays made specifically for blood?

    Read More

    Abcam community

    Verified customer

    Asked on May 24 2013

    Answer

    My guess is that the levels of OXPHOS proteins in whole blood are below the level of detection for activity assays. The activity assays for the most part require higher loading than the sandwich ELISA assays. We have tested Complex IV Quantity with whole blood with good results.



    The caveat is that whole blood must be fresh or must be frozen viable (protocols can be found in the literature). Whole blood should not be loaded in terms of mg of protein as most of the protein present in whole blood is albumin, so using the loading guidelines for tissues to load whole blood will result in loading well below the detection level (OXPHOS proteins are present in leukocytes and platelets only). My suggestion for the customer (if they want to use the quantity dipstick kits) would be to mix 300uL of blood with 900uL of extraction buffer, follow the protocol in regards to the rest of the sample preparation (ice incubation, centrifugation, keeping the supernatant and discarding the pellet). I would advise to the customer to add protease inhibitors to the extraction buffer to prevent degradation of proteins after extraction as granulocytes from blood have high levels of proteases, which will be released once the detergent is added.



    For loading on the dipsticks, the blocking buffer should be mixed with 50uL – 100uL of extracted blood so that the final concentration of blocking is 1X. This mixture can then be used to resuspend the gold conjugated antibody dried at the bottom of the well. Once the gold is resuspended the dipstick can be added to the well and allowed to wick the protein fully. Results will not be perfectly accurate (as loading cannot be done in terms of mg of protein), but will be more qualitative in terms of percent of signal from control mean or relative units. This approach will therefore require you to set up a control sample, which should be a pooled sample from several normal donors (10 – 20 donors would be ideal). From our experience, whole blood will not be as accurate as measuring isolated PBMCs only. But it will allow you to have a rough estimate of the trend.

    The advantage of isolating PBMCs is that they could run both activity and quantity dipsticks/microplates by following the same guidelines of cultured cells.

    Read More

    Abcam Scientific Support

    Answered on May 24 2013

    Question

    zunächst vielen Dank für die ausführlichen und schnellen Infos!!! Ich habe mir inzwischen das Kit ab109908 (Complex II Enzyme Activity Microplate Assay Kit) bestellt und habe jetzt zunächst nur eine wichtige Frage - benötige ich zusätzlich Protease Inhibitoren oder sind diese schon in einem der mitgelieferten Lösungen?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 29 2013

    Answer



    Es werden keinen zusätzlichen Proteinaseinhibitoren für das Kit (ab109908, Complex II Enzyme Activity Microplate Assay Kit) benötigt.

    Wir empfehlen während der Aufbereitung der Proben (bevor das Kit benutzt wird; Proben sollten am Ende 5.5mg/ml in PBS sein) Proteinaseinhibitoren zu benutzen.

    Read More

    Abcam Scientific Support

    Answered on Apr 29 2013

    Question

    Will this kit work with a reader which only reads at 620nm and not the recommended 600nm?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 29 2013

    Answer

    We use DCPIP as a the detector in this kit. DCPIP is an enzyme-catalyzed oxidation electron acceptor that is blue in its oxidized form and colorless in its reduced form. A peak in the absorbance is observed at 600 nm, however you will still be able to read signal at 620nm.

    Read More

    Abcam Scientific Support

    Answered on Mar 29 2013

    Question

    Both kits Complex I and II are working fine.
    However, the same level of activity was measured with incubation buffer only as for the lowest quantity of protein (20µg).
    No activity observed with solution 1 only.

    Question 1 : what is the composition of the incubation buffer? Does it contain a catalyst or a co-factor?

    Question 2 : can the protocol be used by skipping step 6 ? Can the incubation buffer be replaced by another solution?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 13 2013

    Answer

    Question 1 : what is the composition of the incubation buffer? Does it contain a catalyst or a co-factor?

    Incubation buffer is 50 mM Tris.HCl pH7.5, 0.015% lauryl maltoside, and contains 1/20 volume of blocking buffer which is a proprietary casein based blocking buffer. No cofactors, or activators are present that enhance the enzyme activity.

    Question 2 : can the protocol be used by skipping step 6 ? Can the incubation buffer be replaced by another solution?

    For step 6.6 A blocking reagent is in place as a precaution for some samples to prevent significant non specific binding to the plate, but where significant dilution of sample occurs I would be comfortable with eliminating the blocking reagent from incubation buffer or replacing with Blocking solution with 1% BSA in final incubation buffer

    Read More

    Abcam Scientific Support

    Answered on Mar 13 2013

    Question

    I would like to know about the details of the capture antibody of this kit.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 10 2013

    Answer

    Thank you for your inquiry. This antibody was raised against Bovine (cow) complex II. It is cross reactive with human, cow, mouse and rat and is a mouse monoclonal isotype IgG1. It is Western blot negative so we don’t know exactly which subunit it binds to but we know it isolates purified functional complex II. If you have other questions, please contact us.

    Read More

    Abcam Scientific Support

    Answered on Jan 10 2013

    Question

    I have one more question: I need to so this assay under reducing conditions, eg with addition of DTT to the mitochondrial preparation. Would that interfer with the assay for either complex I or II?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 27 2012

    Answer


    We haven't specifically tested the complex I or II kits under reducing conditions and cannot guarantee that it will not affect the enzyme activities. With that being said, my colleagues believe that a reducing agent at moderate concentration shouldn't affect the assay capture step significantly, especially if the reduced samples are sufficiently diluted in the wells.

    Read More

    Abcam Scientific Support

    Answered on Dec 27 2012

    Question

    I was hoping you could also advise me of the conversion formula from mOD/min to nmol/min for both complex I and II activity assays.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 27 2012

    Answer


    To convert from mOD/min to nmol/min you can use the extinction coefficient for the dye in that kit.
    For the complex I kit (ab109721), the dye Epsilon at 450nm = 37 mM-1 cm-1

    The height of 200 uL of liquid in each well is approximately 0.6 cm, therefore epsilon is 22.2 mM-1 well-1 for the complex I kit.

    For the complex II kit (ab109908), the dye Epsilon at 450nm = 21 mM-1 cm-1, which for 200uL of liquid (0.6 cm) equates to 12.6 mM-1 well-1.

    So the change in absorbance per minute in each well divided by 22.2 for complex I or 12.6 for complex II is the mM oxidized per minute.

    Read More

    Abcam Scientific Support

    Answered on Dec 27 2012

    Question

    Inquiry:
    Inquiry: 1. Could a detergent from mitochondrial activity assay kit I (ab109721) or IV (ab109911) be used for the mitochondrial activity assay kit II (ab109908)?
    2. For mitochondrial assay kit II (ab109908) when the activity buffer is added on top of the lipid mixture in the wells is this mixed with the lipid mixture before reading?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 17 2012

    Answer

    Thank you for contacting us and your interest in our products. I am sorry for the delay in getting back to you.

    The three kits mentioned (ab109721, ab109911 and ab109908) use 10% lauryl maltoside detergent to prepare the sample at 5 mg/mL. Therefore this is interchangeable between all three kits.

    In regards to your second question, we find simply adding the buffer to the lipid mixture is enough. There should not be a need for mixing.

    I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Abcam Scientific Support

    Answered on Dec 17 2012

    Question

    I am using the Complex II Enzyme Activity Microplate Assay Kit (ab109908). To detect the activity of SDH in my cancer cells I need to use a higher protein concentration than recommended in the protocol. For that reason, the dilution with incubation solution (step 7-A-1 in the protocol) represents a problem. I have been able to obtain good activity kinetic curves without diluting the samples with incubation solution, using both my cells and abcam's bovine heart mitochondrial lysate. I wonder if this is a correct procedure, i.e. what exactly is the purpose of the incubation solution and can I trust my data if I do not use it - skip step 7-A-1?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 12 2012

    Answer

    Thank you for your inquiry and your patience with this reply.

    The incubation solution contains a blocking buffer to prevent non-specific binding with the antibody. Since your sample extracts are not very concentrated to start with, we recommend to always run a background control with the buffer used to extract the sample (as you will be loading the sample neat). If you prepared the sample according to the protocol, this background control should be 1X detergent in PBS. Also, we recommend to run the samples not just at the highest concentration but at least at 2 or 3 concentrations (1:2 and 1:4) to ensure that the signal is not saturated (diluted in the same buffer as the background control) and that regardless of loading concentration the same result is obtained after taking into account a small dilution factor. It is important for you to ensure that the assay is performed within the linear range of the assay.

    I hope this information helps. Please contact us with any other questions.

    Read More

    Abcam Scientific Support

    Answered on Dec 12 2012

    Question

    Hi

    The products are:

    Complex I Enzyme Activity Microplate Assay Kit (ab109721)

    Complex II Enzyme Activity Microplate Assay Kit (ab109908)

    Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)



    Thanks

    Regards

    Read More

    Abcam community

    Verified customer

    Asked on Dec 07 2012

    Answer

    Thank you for your enquiry.


    I can confirm that In each of these kits we use a non-ionic native detergent at an optimized concentration to maintain OXPHOS complex assembly and activity. This detergent is 1% final N-dodecyl-b-D-maltoside (aka lauryl maltoside). This detergent has been used for many years, particularly in the field of mitochondrial biology and in blue native PAGE separation of OXPHOS complexes.

    I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2012

    Question

    Dear Abcam, My group recently purchased the activity kit "ab109908", for measuring ETC complex II activity. After some initial problems using the kit for our mouse breast cancer cell lines (had to use a higher protein concentration than recommended in the protocol), we finally got it to work. The problem is that the kit (at least the one we got) did not provide enough ubiquinone2 to run a full 96-well plate - today I ran the last two strips and needed 10ul but there were only 2.6ul left! A still reading a signal but cannot compare to the previous experiments, which was the all purpose of this run to have statistical significance. Would it be possible to receive an extra kit so that we can complete our study, or at least part of the kit that allows to carry these final runs. Please let me know, as we need these data ASAP.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 28 2012

    Answer

    Thank you for contacting us. I am sorry to hear that the full amount of Ubiquinone 2 was not present in this kit and apologize for any inconvenience this has caused. Provided that this purchase has been made within the last six months, I would be very happy to provide a free of charge replacement to you. In order to process this could you please provide the Abcam order confirmation number or the PO and date of purchase. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Abcam Scientific Support

    Answered on Nov 28 2012

    Question

    To whom it may concern with,

    What kind of the detergent is in the Complex Enzyme Activity Microplate Assay kit from MitoScience? Also I'd like to know the concentration of it. Because I want to use the same detergent for BN-PAGE. Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 24 2012

    Answer

    Thank you for your inquiry.
    The 10xdetergent of this kit is Lauryl maltoside 10%.
    Thank you.

    Read More

    Abcam Scientific Support

    Answered on Oct 24 2012

    Question

    Thank you very much for the prompt response and the information on complex I activity.

    I also use the following kits:

    ab109908 (complex II enzyme activity),

    ab109909 (complex IV activity),

    ab109716 (ATP synthase activity) and

    ab119692 (citrate synthase activity).

    Could you please as well let me know which factors should I use to convert them to mM/min instead of mOD/min? I use and will use often these kits for a large study.

    Thanks in advance for your great help and best regards

    Read More

    Abcam community

    Verified customer

    Asked on Oct 05 2012

    Answer

    Thank you for contacting us.

    Same process for converting all to molarity per amount of protein loaded into well.
    Note these are all per cm and should be converted into path length for microplate which should be about 0.6cm.

    Extinction coefficients

    ab109908 (complex II enzyme activity),
    ε600 = 21 mM-1cm-1

    ab109909 (complex IV activity),
    ε550 = 21.1 mM-1cm-1

    ab109716 (ATP synthase activity)
    ε340 = 6.220 mM-1cm-1

    ab119692 (citrate synthase activity).
    Ε412 = 14.1 mM-1cm-1

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Oct 05 2012

    Question

    Initially I tried one hour, then two hours. After that, I incubated overnight, but didn't see much difference in absorbance. I'd appreciate trying a new kit.The po # is

    Read More

    Abcam community

    Verified customer

    Asked on Sep 21 2012

    Answer

    Thank you for your reply.

    I will be happy to send out a new kit for you to try. I think I have found your original order, but I just want to double check the address before having the new one sent. Was the original placed on **shipped to:

    **

    If this is the correct information, then please let me know and I will have the new kit shipped out for delivery next week.

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Sep 21 2012

    Question

    Thank you for your response.
    1. The mitochondrial lysate is below:
    Mouse Brain Mitochondrial lysate
    (Abcam # ab110351)
    2. I am using lysates from cell lines that I prepared as described in the kit's manual.
    3. The samples start off with signal which doesn't decrease (as would be expected) over time.
    4. I can certainly send you results (but am away until tomorrow). In any case, there is not much to show. As noted, there is signal at the start of the assay but these doesn't decline as would be expected.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 20 2012

    Answer

    Thank you for your reply.

    I just had one further question, you say that you start off with signal, but it does not decrease with time, as it should. I was curious how long you kept measuring for? I only ask, as some samples take a little longer to start producing signal than others.

    By the sounds of it though, I think the best way forward is to go ahead and send out a new kit for you. To do this you could provide either the Abcam order number or the PO# used to purchase the kit?

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Sep 21 2012

    Question

    Unfortunately, I have not been able to get this kit to work. I ordered your mitochondrial control lysates, but I see no activity with these either.
    Would you suggest anything else to try? Perhaps the kit is defective?
    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 19 2012

    Answer

    Thank you for contacting Abcam.

    I am sorry about the issues you are having with ab109908, if we cannot resolve the issues you are having then I would be more than happy to send a new kit.

    I just have a few questions for you:

    1 - What is the catalogue number of the mitochondrial lysate you are using?

    2 - What type of samples are you testing?

    3 - Is there any signal coming from the kit, or are all wells just giving a '0' reading?

    4 - Would it be possible to send a copy of the results that you are getting with this kit, so that I can talk to the lab about this?

    I look forward to your reply and helping to resolve this issue?

    Read More

    Abcam Scientific Support

    Answered on Sep 19 2012

    Question

    In sample prep section:
    kit for complex I: "spin 12,000 g 20min, no temp given"
    kit for complex II: "spin 25,000 g 20min, 4 degree given"
    1) can samples prepared from each kit be spun at same speed and Temp?
    2) if so, which speed and temp?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 08 2012

    Answer

    Thank you for contacting us.

    The lab let me know that +4C at either speed would befine for both kits.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Sep 08 2012

    Question

    I am using ab109721 and ab109908. They are very similar kits and both contain a 20x Buffer. Is the 20x Buffer from both kits the same buffer?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 31 2012

    Answer

    These solutions are not the same… different buffers and 10-fold different detergent concentrations.

    I hope this is helpful. Please contact us again if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Aug 31 2012

    Question

    An olderprotocol said the kinetic assay could be read at 450 nm. The protocol on Abcam's website only says 600 nm. Was the 450 nm a typo? Is there a way to alter the protocol and measure at 450 nm? The customer can only read at 450 and 700 nm.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 28 2012

    Answer

    Thank you very much for your call yesterday and for your patience while I'vebeen in touch with the labregarding your enquiry.

    We couldn't find a record of the kit being measured at 450 nm or of previous instructions in the protocol to measure at 450 nm. 600 nm is the optimum absorbance for this particular dye (highest absorbance). But these dyes have pretty wide absorbance ranges (shoulders on the curve) so it's likely the decrease in color could be measured at wavelengths other than 600, butwe can't say for sure. At the very least, the next time we measure it we will record the 450nm spectrum in addition to the 600.

    I hope that this information will be useful, but if you have any further questions or concerns please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Aug 28 2012

    Question

    step 6. Can use 16,000g for a longer time instead of 25'000g for 20 min?
    step 3. Can dilute sample in 1x incubation solution instead of PBS, becuaseneeds to use 250 ug/well for the assay and cannot dilute further after the detergent step (step 4).

    Read More

    Abcam community

    Verified customer

    Asked on Aug 10 2012

    Answer

    Thank you for contacting us.

    The lab already got back to me:

    step 6. Can use 16,000g for a longer time instead of 25'000g for 20 min?
    Answer: Yes, same time - 20 min


    step 3. Can dilute sample in 1x incubation solution instead of PBS, becuaseneeds to use 250 ug/well for the assay and cannot dilute further after the detergent step (step 4).
    Answer: USE PBS, EXTRACT THE SAMPLE WITH DETERGETNT. FINAL CONCENTRATION IS 5 mg/mL ADD 50 uL TO THE WELL, SO TOTAL AMOUNT 250 ug, DON’T USE INCUBATION SOLUTION AT ALL.



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 10 2012

    Question

    Thank you for your answer.

    So i would like you to check if the integration of the two protocols i've done is correct:





    1. After harvesting the cells, i resuspend pellet in PBS to an optimal concentration of at least 10million cell\ml and froze -20C prior to extraction (As said in page four of the protocol you've send me)

    2. 50ul of cell pellet+400ul PBS+ proteases inhibitors (can i use the same inhibitors i currently use for protein extraction for western blotting, such asAprotinin, Leupeptin, PMSF?)

    3. Add 50ul of lauryl maltoside 15% to 450 ul of the previous cell-solution

    4.mix well and incubate on ice 30min

    5.16.000xg 20min 4C

    6. collect supernatant and quantify protein concentration with BCA method

    7. Resuspend pellet to 5.5mg\ml in PBS

    8. Add 1/10 volume of Detergent to the sample (from the kit Complex II Enzyme Activity Microplate Assay Kit)

    9. mix well and incubate on ice 30min

    5. 25.000xg 20min 4C

    6. collect supernatant


    7. Dilute sample in 1X incubation solution to 1.2 mg/ml(prevously prepared as said in the User protocol)

    8.Add 50ul of diluted sample in each well. Incubate 2hour at room temperature

    9. procede with the Measurement protocol

    Read More

    Abcam community

    Verified customer

    Asked on Jul 04 2012

    Answer

    Thank you for your reply.

    I am sorry for the misunderstanding.

    The samples preparation guide is not in addition to the kit protocol but an explanation of the protocol. These arethe steps you should take:

    1. Resuspend sample – cell culture pellet (ca 50ul)in 450ulPBS.

    2. Add 1/10 volume of Detergent to the sample (e.g. if the

    total sample volume is 500 μl, add 50 μl of Detergent).

    3. Mix immediately and incubate for 30 minutes on ice.

    4. Centrifuge at 25,000 g for 20 minutes at 4°C. Collect the

    supernatant discard the pellet.

    5. Collect supernatant and quantify protein concentration with BCA method. Make sure the protein concentration is 5 mg/ml. Dilute in incubation solution if necessary.

    Now continue with the protocol.

    I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 04 2012

    Question

    Our customer purchased ab109908 and ab109909 and received them last Monday.
    Since that day, the customer stored whole kits in deep freezer for about 10 days.
    Could you let me know if there's simple test method to check those kits work or not.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 04 2012

    Answer


    I am afraid the components Tube 1, detergent, and microplateofab109909 may be damagedas they are not supposed to be frozen at all.
    The components of ab109908seem to be a bit more robust.
    Unfortunately, there is no quick test to check the activity of the kits.


    I hope this information is still helpful to your customer.

    Read More

    Abcam Scientific Support

    Answered on Jul 04 2012

    Question

    Does kit allow fluorescence detetcion or can a fluorescent probe be used with it?
    Alternatively, do we have fluorometric kit for measuring complex II activity?

    Read More

    Abcam community

    Verified customer

    Asked on May 29 2012

    Answer

    Thank you for contacting us.

    The lab let me know the following:

    The question is really interesting. Unfortunately without more testing the answer is no.

    We use DCPIP (http://en.wikipedia.org/wiki/Dichlorophenolindophenol) as a the detector in this kit which is not fluorescent. We are not aware of any fluorescent redox dyes that would accomplish this function or a way in which we could couple the reaction.

    But we can certainly follow up on it. We are not aware of any other kits like this.

    I hope this information is of some help to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on May 29 2012

    Question

    Hi, I have recently placed an order for the following kits:

    Ab109721

    Ab109875

    Ab110172

    Ab109911

    Ab109908

    Would you be able to let me know if these kits have been successfully used with previously snap-frozen tissue that has been stored at -80C? I am waiting on fresh mouse heart tissue, but am wondering if I could also use these kits to assay tissue I already have stored.

    Many thanks.

    Best wishes,

    Read More

    Abcam community

    Verified customer

    Asked on May 23 2012

    Answer

    Thank you for contacting us.
    These kits will be OK to use with snap frozen tissues.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on May 23 2012

    Question

    I'm trying to optimize this kit with tissue samples. The signal is very low, close to non-detectable. Positive control is mitochondrial extracts from cells and itsreadings are good, so the assay is working.
    How can I increase the signal in the tissue samples? Is there a way to concentrate the sample from the very beginning? For example, I'd like to concentrate the sample more than 5.5 mg/mL in PBS. Will I then need to add proportionally more detergent to this sample? If I concentrate the sample to 10 mg/mL, do I have to use double the detergent? Or can more than 50 uL of sample be added to the each well?
    Any suggestions would be welcome. Thank you!

    Read More

    Abcam community

    Verified customer

    Asked on May 22 2012

    Answer

    Thank you for your call last week and for your patience while I've been in touch with my colleagues at MitoSciences in regards to your questions.

    I've heard back from one of the lab scientists, and following are her suggestions:

    "Increasing the homogenate protein to 15 – 25mg/mL may help.My suggestion is to test higher homogenate concentration solubilized with 1% lauryl maltoside as well as 0.5% lauryl maltoside and test them on the plate side by side. Note that the detergent that comes with the kit is at 10% (so 1/10 dilution gives 1% detergent).

    The standard concentration for solubilizing proteins for an activity assay is 1% lauryl maltoside. In our experience more detergent is not always best (particularly for complex V and PDH). An example is that a membrane enriched fraction of rat liver homogenate gives a higher signal when the sample is at 25mg/mL than when it is at 5mg/mL, similarly this sample gives a more linear activity when it is solubilized at 0.25% lauryl maltoside than at 0.5% or even 1%.Not all the samples behave in the same way for complex V. Whether this experience will help fix the problem in complex II, I don’t know but it is worth a try.

    Another thing you could do is to generate a crude enrichment of membrane proteins before addition of detergent.This can be accomplished by centrifuging the homogenate at 25,000 g for 10 minutes at 4C, discarding the supernatant and resuspending the pellet in half of the original volume in the solution buffer of the assay.This can be followed by protein measurement and addition of the assay detergent. After addition of assay detergent the sample will be the supernatant and the pellet can be discarded.

    We have seen that this membrane enrichment is crucial for complex V activity assay, and although we have not tested it in complex II assay I predict it may help to increase the signal."

    I hope this informaiton will be useful, but if you have any further questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on May 22 2012

    Question

    Should the OD be read every 20 seconds or every 1 minute? Both are written in the protocol.

    Read More

    Abcam community

    Verified customer

    Asked on May 09 2012

    Answer

    Thank you for your call today and for your patience while I've been in touch with my colleagues at MitoSciences regarding your enquiry.

    The OD can be read either every 20 seconds or every 1 minute, whichever is easiest for you to do. In our lab, the OD is measured every 20 seconds, but 1 minute intervals will also give plenty of data.

    I hope this information will be helpful, and I apologize for the confusing instructions. If you have further questions or need anything else, please don't hesitate to ask.

    Read More

    Abcam Scientific Support

    Answered on May 09 2012

    Question

    I would like to use human planctal tissue extract with this kit. Do you have any recommended protocols for extraction of this sort of tissue with
    Complex II Enzyme Activity Microplate Assay Kit (ab109908)?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 19 2012

    Answer

    We have not worked with this kind of tissue yet. We do have a protocol with general principals of cultured cell and tissue preparation I hope will be useful:


    http://www.mitosciences.com/PDF/mitosciences-sample-preparation-guide.pdf
    I hope thishelps. Please contact us again if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Apr 19 2012

    Question

    Are the blocking buffers in the complex II kit ab109908 the same as in the PDH combo kit ab110671? Is the buffer ab126587?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 17 2012

    Answer

    Thank you for your call today and for your patience while I have been in touch with the lab about these kits.

    I can confirm that the 10x blocking buffers are the same in each kit, soit will be fineto interchange them. This buffer is available for purchase separately as ab126587 as well.

    I hope this information will be useful, but please let me know if you have further questions or if there is anything else that we can do for you, and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Apr 17 2012

    Question

    Thank you very much!



    The amount is not a problem...I can have as many flies as I want!



    I think I would like to try the kit....BUT how can I buy the kit without knowing if I can use tecan reader?!

    Any idea? My boss wont approve buying it....without knowing we have the machine to read it:) LOL



    Bw

    Read More

    Abcam community

    Verified customer

    Asked on Mar 23 2012

    Answer

    Thank you for your reply. My colleague Cathrin is not in the office today.

    Unfortunately, I do also not know if this kit is compatible with a Tecan reader.

    I can suggest to ask the manufacturer of the Tecan reader. They should be able to help you after they had a look at the datasheet of protocol of this kit.

    I am sorry I could not be of more help and hope this information is nevertheless helpful.

    Read More

    Abcam Scientific Support

    Answered on Mar 23 2012

    Question

    Do theywork on mice andDrosophilla? How specific is it? Tecan machine compatable (Flourescence)?Does the last buffer in the protocolincludes or need a detergent?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 22 2012

    Answer

    Thank you for contacting us.

    I have some good and some bad news for you. the good ones are that it will work in mice, and that the kits may work with Drosophila samples as we as the homology is for both enzymes compared with the human proteinsis over 98%. The bad one is that you will need quite an amount of material to get your experience running, and this may simply not possible with this test species.

    These assays are specific to these enzymes and isoforms as demonstrated by mass spec analysis of immunoprecipitates.


    Regarding the buffers, adetergent is not included in either final buffer.

    And last but not least, unfortunately the lab couldn't tell me if your "Tecan reader" will work with the kits: they don't know it.



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Mar 22 2012

    Question

    Our customer has again tested and her results were unsuccessful.

    See attached her graph and her correspondence below;

    I have tried out the protocol again, without success. Please find attached a copy of my graph from the recent experiment, show not much reduction in OD.

    I wondered if you could point me in the direction of someone, preferably in Australaia, who have tried this kit with success, so that I can talk to them about sample preparation?
    Any further advice from the company will be also much appreciated.

    Any other suggestions would be great.

    Looking forward to your response.

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 16 2012

    Answer

    Thank you for your reply which has been forwarded to me as my colleagueis currently away from the office.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad kit.

    I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

    Regrettably, we are not able to provide information regarding contactdetails of other customersas this is confidential.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on Mar 16 2012

    Question

    Our customer has responded with the following questions;



    I did a trial using Hela cells, but there was no SDH activity at all. I think the problem is probably in the sample preparation, and I would be grateful if you or the company could assist me in clarifying a few points:



    1. What is the optimal way to prepare the sample to add to the detergent?

    In the protocol, it specifies that it should be at a protein concentration of 5.5mg/mL- should the sample be lysed in a particular method and quantified prior to loading?

    How would I be able to quantify the sample if it is a cell pellet that I am using in the first step?

    Should the cell pellet be homogenised prior to adding to detergent?

    The protocol specifies that the tissue homogenate or cell pellet should be re-suspended in PBS.





    2. In using the incubation solution, what volume should be used?
    For example, using a Hela cell pellet tobegin with,if at the end of the sample preparation, I have 50uL of 5mg/mL protein plus detergent, is it right that I add 12uL of this to 38uL of incubation solution to bring it to 60ug in 50uL?



    3. At what temperature should the lipid solution be incubated? Room temp or 4degrees?



    Looking forward to your response.





    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 23 2012

    Answer

    Thank you for your inquiry.

    1.)

    Please follow this link to apreparation guidewith some suggestions for sample preparation.

    http://www.mitosciences.com/PDF/mitosciences-sample-preparation-guide.pdf

    I have copied and pasted the relevant section below for you. The trick is to get the sample to approximately 5 mg/mL during extraction.

    Why do we do this – well this prevents a huge detergent:enzyme ratio which could destroy the enzyme or conversely having too much protein to detergent and getting a poor extraction efficiency.

    Protein Extraction Method II with 10X Lauryl Maltoside:

    1. Resuspend and dilute the cell pellet with 8 volumes of PBS (e.g. 50 μL pellet + 400 μL PBS to a total volume of 450 μL). Add protease inhibitors.
    2. Add a 10-fold dilution of 10X lauryl maltoside (e.g. add 50 μL to the cell suspension for a total volume of 500 μL).
    3. Mix well and incubate on ice for 30 min.
    4. Centrifuge at 16,000 x g for 20 min at 4oC.
    5. Collect supernatant and discard the pellet.
    6. Measure protein concentration with BCA™ Assay or NanoDrop™ Spectrophotometer.

    The protein concentration of the extract should now be ≤ 5 mg/mL, depending on extraction efficiency, as measured by BCA™ Assay. To measure protein concentration with a NanoDrop™ Spectrophotometer, follow the manufacturer’s instructions for measuring protein. In the pull-down menu for sample type, select “Other protein (E 1%),” then enter an extinction coefficient (Ext. Coeff, E 1% L/gm-cm) from Table 3 below.

    Table 3: Extinction Coefficients for Cell Extracts

    HepG2 16.2

    HeLa 14.4

    HL-60 17.2

    It may be possible to freeze the extract at -80oC before running the assay without significant loss of activity.

    2.)I agree with your customer, this looks right.

    3.)The solution should be incubated at roomtemperature.

    I hope this information is helpful and wish your customer good luck with their experiments.

    Read More

    Abcam Scientific Support

    Answered on Feb 23 2012

    Question

    Hi Technical,



    I hope you are well J



    Please see below our customer correspondence;



    Thank you for helping me with this kit. Here are my questions:



    1. What sequence does the complex II antibody bind to?

    2. What is the optimal method for mitochondrial preparation? I am using a cell line, and have a protocol in the laboratory for mitochondrial fraction enrichment. Is that sufficient?

    3. What concentration of mitochondrial concentrate is necessary? The protocol details recommended concentrations for whole cell extract or tissue mitochondria only.

    4. Could you tell me the components for the reagents? This is to allow me to better understand how the reactions work- buffer, detergent, blocking solution, activity buffer, lipid mix.

    5. Is the microplate strips compatable with the Veritas Microplate Luminometer?



    Any help would be greatly appreciated J



    Looking forward to your response.





    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 17 2012

    Answer

    Thank you for your inquiry. Please find the answers to your customer's questions below:

    1. What sequence does the complex II antibody bind to?

    Unfortunately this is unknown, due to requirement for native binder we immunize with native enzyme so binding site is very hard to determine.

    2. What is the optimal method for mitochondrial preparation? I am using a cell line, and have a protocol in the laboratory for mitochondrial fraction enrichment. Is that sufficient?

    Most of our protocols have been developed without the need for mitochondrial isolation therefore we quote amounts of tissue or cell lysates. If you wish to use mitochondria our experience is there is only around a 3X enrichment but this will make the sample more active per mg.

    And therefore will likely work well .

    3. What concentration of mitochondrial concentrate is necessary? The protocol details recommended concentrations for whole cell extract or tissue mitochondria only.

    As with cell culture lysates I would recommend 4-250ug/well

    4. Could you tell me the components for the reagents? This is to allow me to better understand how the reactions work- buffer, detergent, blocking solution, activity buffer, lipid mix.



    Detergent : Lauryl maltoside 10%

    Concentrated Wash Buffer: 0.4M KPi pH7.2, 0.3% LM

    Activity Buffer: 20 mM KPi, 0.015% LM

    Concentrated Succinate: 1M Sodium succinate.hexahydrate

    Concentrated DCPIP : 2.5 mM DCPIP

    Concentrated Ubiquinone: 13 mM UQ2

    Blocking buffer: Casein based proprietary blocking buffer

    Lipids : proprietary mix of phosphatidyl choline


    5. Is the microplate strips compatable with the Veritas Microplate Luminometer?

    No, I do not believe it is. Veritas is a luminometer – it measures light emission only. Required is a standard laboratory absorbance microplate reader.

    http://en.wikipedia.org/wiki/Plate_reader

    I hope this information is helpful and wish your customer good luck with their research.





    Read More

    Abcam Scientific Support

    Answered on Feb 20 2012

    Question

    uses human skeletal muscle biopsies from children (i.e. has limited samples)
    How much tissue do I need to startwith?
    Is this different for each kit?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 02 2012

    Answer

    Thank you for contacting us.

    I have heard back from the lab with the following information:

    All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/microplate well.
    The good news is that normal whole muscle homogenate contains a good robust mitochondria signal so less is required, muscle samples are described in most if not all protocols.
    Another good news is that the same preparation for any of the assays is suitable for all others. I would estimate that if a 1 mg of homogenate can be prepared (0.2mL at 5mg/mL).
    This would provide enough material for each of the assays when diluted to the specific concentration in each assay.
    We would recommend - if contemplating doing multiple assays - to include a mitochondrial sample as positive control such as ab110338 isolated mitochondria (note this is cow source)



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Feb 02 2012

    Question

    Dear technical, Good day! We have a customer who is interested in the above kit and has the following technical enquiries regarding: 1. What sequence does the complex II antibody bind to? 2. What is the optimal method for mitochondrial preparation?  I am using a cell line, and have a protocol in the laboratory for mitochondrial fraction enrichment.  Is that sufficient?  3. What concentration of mitochondrial concentrate is necessary?  The protocol details recommended concentrations for whole cell extract or tissue mitochondria only. 4. Could you tell me the components for the reagents?  This is to allow me to better understand how the reactions work- buffer, detergent, blocking solution, activity buffer, lipid mix. 5. Is the microplate strips compatable with the Veritas Microplate Luminometer? I hope to hear from you soon. Kind regards    

    Read More

    Abcam community

    Verified customer

    Asked on Nov 30 2011

    Answer

    Thank you for contacting us. Please find below the answer to your questions. 1. What sequence does the complex II antibody bind to? This antibody was generated by protein immunization and not by peptide immunization and therefore we don’t know the exact epitope it binds 2. What is the optimal method for mitochondrial preparation? I am using a cell line, and have a protocol in the laboratory for mitochondrial fraction enrichment. Is that sufficient? You don’t need to isolate mitochondria from cells to use this assay. However if you want to use the kit with mitochondria preparation from cells, we suggest to use Mitochondria Isolation Kit for Cultured Cells designed to isolate mitochondria from cultured cells: https://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-ab110170.html 3. What concentration of mitochondrial concentrate is necessary? The protocol details recommended concentrations for whole cell extract or tissue mitochondria only. Optimal loading for samples not specified in the protocol, will have to be determine empirically by the user. 4. Could you tell me the components for the reagents? This is to allow me to better understand how the reactions work- buffer, detergent, blocking solution, activity buffer, lipid mix. The detergent is Lauryl maltoside (which is also available from the catalogue as a standalone product). The buffer is a phosphate based buffer and the activity buffer contains a ubiquinone, succinate and DCPIP (2,6-diclorophenolindophenol). The reaction is as follows: Succinate + ubiquinone (Q) ------- Fumarate + ubiquinol (QH2) ubiquinol (QH2) + DCPIP ------- ubiquinone (Q) + DCPIPH2 5. Is the microplate strips compatible with the Veritas Microplate Luminometer? It is necessary to have a spectrophotometer plate reader capable of measuring absorbance at 600nm. The complex II reaction does not emit light and cannot be read by luminometry. I hope this information is helpful. Please do not hesitate to contact us if you need further advice or information.

    Read More

    Abcam Scientific Support

    Answered on Nov 30 2011

    Question

    There is no reduction in optical density with our samples.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 29 2011

    Answer

    Thank you for bringing this to our attention. An easy way of determining whether the kit is working or not is to use one of the positive controls as shown in the kit protocol (HepG2 lysate, Bovine heart mitochondria, Rat heart mitochondria). BHM and RHM are available from the catalogue. Loading at 250ug/well (2.5ug/uL) should yield -3mOD/min https://www.abcam.com/Bovine-Heart-Mitochondrial-lysate-ab110338.html https://www.abcam.com/Rat-Heart-Mitochondrial-lysate-ab110347.html Protease inhibitors do not affect the assay. It is also possible that Lung cells have a very small amount of complex II (the stochiometry of ETC complexes is not 1:1, so if you get activity with complex I it does not mean you should have a similar activity of complex 2 in the cell line). If this is the case, you may need to isolate mitochondria from the cell line. An option to do this is the following kit: https://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-ab110170.html" If you have used all the reagents, I will be happy to send another kit, but I do recommend including the controls, given the possibility that the samples are deficient in Complex II compared to Complex I.

    Read More

    Abcam Scientific Support

    Answered on Nov 29 2011

    Question

    Kits were stored at room temperature for over 2 weeks and are probably no longer viable.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2011

    Answer

    Thanks for your call today and letting us know about the problem with the storage instructions on our shipping boxes. As we discussed, we are sending free of charge kits ab109908 and ab110671 on the order ***, which should arrive tomorrow. I apologize for all the trouble and stress with this issue, and I wish you the best of the luck with your future endeavors. Please let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2011

    Question

    Can the samples be forzen down after step A-6?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 14 2011

    Answer

    I have talked to the lab as to whether you can freeze the samples after step 6 and they have told me that it should be ok to freeze the samples down and then use them the next day. You just have to careful and also make sure that you are comparing frozen to frozen samples and not fresh to frozen.

    Read More

    Abcam Scientific Support

    Answered on Nov 14 2011

    Question

    Thank you very much for your reply, I forwarded the mail to the customer. She now explains her project and wants me to ask you if you have any ideas, or products which she might be able to use.   Here is her mail:   Thanks so much for finding out from the company for me. I have rodent mitochondria samples that I would like to use, and preferably I would like to test all 4 or 5 of the electron transport chain complexes - that is why I mentioned the 5x96-well plate assay as it seemed to me that this could test all 5 complexes. The reason I am interested in this product is to find an alternative to the Oxygraph method that measures oxygen consumption and electron transport chain complex functioning. I am however at the stage where I do not know whether all 5 of the complexes' activity will be altered with my intervention (an antiretroviral drug). I have looked through the database and discussed this with my promotor, and we suspect that as a first step we could try the MitoProfile Membrane Integrity Cocktail antibody as well as the ATPase/Complex V specific activity to get an idea of what is happening. I have previously ordered the MitoOxphos Complex I-V Cocktail (and will order it again soon - I require more), and I believe that this might give us better leverage as how to go forward with our hypothesis. My hypothesis is that one of the antiretroviral agents used in HIV/AIDS treatment can induce mitochondrial abnormalities, and studies have shown various effects ranging from changes to mitochondrial DNA, mitochondrial gene expression and membrane integrity. However my study is quite novel and we are unsure whether our experimental conditions would confirm previous findings, and if the drug could alter not only gene and protein expression levels of the complexes, but affect functioning with ATP production and structural integrity. You are welcome to pass this information on to your contact at Abcam - perhaps with this info they can advise more appropriately. I do however think that the ATPase specific activity assay might be a very good choice to later substantiate Oxygraph analyses.  

    Read More

    Abcam community

    Verified customer

    Asked on Nov 10 2011

    Answer

    Thank you for your reply. I have been in touch with our colleagues at Mitosciences who have kindly provided the following information: To answer your question directly, The antibody coating the plate supplied with Kit MTOX1 ab109903 does not cross-react with rat samples (it is human/bovine only).  If you would like to  test complex I activity in Rat mitochondria, it would be better to purchase kit ab109121.  Click here (or use the following: https://www.abcam.com/Transglutaminase-2-antibody-EPR2956-ab109121.html). However, the drawback of kit ab109121 is that it is not rotenone sensitive as it is only the dehydrogenase activity of the enzyme and does not follow the complete reaction (dehydrogenase-ubiquinol reductase).  This is explained in detail in the protocol on the datasheet. To answer the complete question, we have had a substantial experience testing mitochondrial toxicity due to antiretrovirals.  It looks to me that you are treating animals with an antiretroviral compound for HIV?  The effect on mitochondria due to antiretroviral therapy varies tremendously depending on the regime used and to my knowledge the most prominent causes of mito toxicity are : (1) inhibition of chain elongation and/or exonuclease activity of the mtDNA polymerase γ, (2) inhibition of thymidine kinases, (3) oxidative stress, (4) impairment of the adenine nucleotide translocase and (5) reduction in mitochondrial membrane potential (Δψm).  The easiest and most economical way of looking at such a wide spread of potential effects in mitochondria is by testing cultured cells exposed to the compound with the following sets of kits: ab113849 ATP luminescent detection kit = check the scientific support tab as well as the protocol for further insight into mitochondrial toxicity testing.  This can test the mitochondrial bioenergetic state of the cell by comparing treatment in glucose vs. galactose based media. Click here (or use the following: https://www.abcam.com/Luminescent-ATP-Detection-Assay-Kit-ab113849.html). ab113850 JC1 or ab113852 TMRE.  Both of these test membrane potential.  The protocol also has insights as to the effect of low membrane potential in the overall mitochondrial metabolism. Click here (or use the following: https://www.abcam.com/JC-1-Mitochondrial-Membrane-Potential-Assay-Kit-ab113850.html). Click here (or use the following: https://www.abcam.com/TMRE-Mitochondrial-Membrane-Potential-Assay-Kit-ab113852.html). ab113851 DCFDA = which measures ROS. Click here (or use the following: https://www.abcam.com/DCFDA--H2DCFDA-Cellular-ROS-Assay-Kit-ab113851.html). ab110217 and ab110216 Mitobiogenesis In-cell ELISA kits (colorimetric or IR) = These will measure the effect of a compound on the protein levels of a mitochondrial DNA encoded protein (typically affected by nucleoside reverse transcriptase inhibitors) in comparison to the protein levels of a nuclear DNA encoded protein (not affected by NRTIs).  Click here (or use the following: https://www.abcam.com/MitoBiogenesistrade-In-Cell-ELISA-Kit-Colorimetric-ab110217.html). Click here (or use the following: https://www.abcam.com/MitoBiogenesistrade-In-Cell-ELISA-Kit-IR-ab110216.html). The kits above mention can be used with any mammalian cell.  The mitobiogenesis kits can be used with human, rat, mouse or bovine cells.  The protocols explain in detail how to set the experiments.  If you haven’t tested the compound with the first basic acute tests on cells, I would recommend to do this.  It will help to narrow down the potential cause of toxicity or at least help have a viable hypothesis of what could be happening in the animal.  If the first four are normal (after acute treatment) I would suggest for to move ahead with the mitobiogenesis kit after treating the cells for at least 5 – 7 passages.  If they have a very good reason to suspect (due to the chemical structure of the compound) that the effect will be only on the mtDNA polymerase, then my suggestion is to go directly to the ab110217 to confirm the hypothesis. Once you have a reasonable hypothesis of the effect of the compound in-vitro, then choosing the test for in-vivo testing will be much easier.  i.e if the retroviral has an important effect on ROS production then they may want to follow up with antibodies that target ROS induced post-translational effects such as anti-nitrotyrosine, ab110282. Click here (or use the following: https://www.abcam.com/3-Nitrotyrosine-antibody-7A12AF6-ab110282.html). If galactose sensitizes the cells to compound toxicity as measured by the ATP luminescent assay, then very likely the compound is affecting the enzyme activity/assembly/levels of one of the complexes of the electron transport chain.  In this instance, then they will need to determine if it is a direct or indirect effect on the enzymes.  The MTOXC kits (5 kits) will test  whether the compound affects directly any of the enzymes using bovine mitochondria (Note that if a compound directly inhibits the transfer of electrons, by in-vitro testing, in bovine mitochondria, it will very likely affect the transfer in of electrons in other mammalian species such as rat).  MTOXC will test activity in the presence of compound (while the assay runs) The following MS kits: Complex I Enzyme Activity Microplate Assay Kit ab109721, Complex II Enzyme Activity Microplate Assay Kit ab109908, Complex IV Rodent Enzyme Activity Microplate Assay Kit ab109911 and ATP synthase Enzyme Activity Microplate Assay Kit ab109714,  all of which are rat reactive) will test indirect effect of the compound in the activity of enzymes (post-translational modifications, assembly defects) in tissues of treated animals.   ab109721 Click here (or use the following: https://www.abcam.com/Complex-I-Enzyme-Activity-Microplate-Assay-Kit-Colorimetric-ab109721.html). ab109908 Click here (or use the following: https://www.abcam.com/Complex-II-Enzyme-Activity-Microplate-Assay-Kit-ab109908.html). ab109911 Click here (or use the following: https://www.abcam.com/Complex-IV-Rodent-Enzyme-Activity-Microplate-Assay-Kit-ab109911.html). ab109714 Click here (or use the following: https://www.abcam.com/ATP-synthase-Enzyme-Activity-Microplate-Assay-Kit-ab109714.html). These MS kits test activity of tissues or cell extracts after they have been exposed to an experimental condition.  For example if you test NRTI compounds with MTOXC, all results will be normal.  However if you test with the MS kits tissues from animals treated with NRTIs for a prolonged period of time, you will see decrease in activity.  You could use the following antibody cocktails: ab110413 MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail  Click here (or use the following: https://www.abcam.com/Total-OXPHOS-Rodent-WB-Antibody-Cocktail-ab110413.html). ab110414 MitoProfile® Membrane Integrity WB Antibody Cocktail  Click here (or use the following: https://www.abcam.com/Membrane-Integrity-WB-Antibody-Cocktail-ab110414.html). If you believe that the structural membrane integrity of the mitochondria (of treated animals) can have an important impact in complexes assembly and protein levels in specific compartments, then these cocktails may be a good choice.  Note that the ATP synthase Enzyme Activity kit ab109714 measures the reverse reactions (ATP hydrolysis) and not ATP synthesis, however the ATP hydrolysis is still oligomycin sensitive.  A decrease in oxygen consumption by the oxygraph could be due to effects in any of the complexes and not just ATPase.  I.e. rotenone affects oxygraph results, but it is a complex I specific inhibitor. So choosing this early one MS541 kit may lead them into a wild goose chase.  You would need to look at mitochondria in a more holistic way and not just as a measure of ATP synthesis.   I hope this information will be helpful to you. if you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Nov 10 2011

    Question

    I am very sorry for this mistake. This is the correct PO: xxxx

    Read More

    Abcam community

    Verified customer

    Asked on Nov 29 2012

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx. As product may become trapped in the caps of our vials, please remember to spin down any vials before opening. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Nov 29 2012

    Question

    What is the guarantee period for these kits?

    Read More

    Abcam community

    Verified customer

    Asked on May 18 2012

    Answer

    Thank you for contacting us.

    The kits are guaranteed for 6 months if stored as recommended on the datasheet.

    Please note there are components in the kits that are supposed to stored at different temperatures e.g. -20 or -80C, so we recommend carefully checking the datasheet and protocol booklet.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on May 18 2012

    Question

    Hi Kate,

    Our #### was dated #####.

    We ordered it through Abcam.

    Let me know if you need any further info.

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 29 2012

    Answer

    Thank you for confirming this information and for your help and cooperation with this case.

    As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

    Credit ID: #####

    As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

    I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

    Read More

    Abcam Scientific Support

    Answered on Mar 29 2012

    Question

    Thanks for replying so promptly, and also for you kind offer to replace our kit. The PO number was xxxxxx. Please let me know if you need any additional information.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 29 2012

    Answer

    Thank you for your reply. Unfortunately the PO information which you have given me corresponds to Abcam Order #xxxxx for ab110338 Bovine Heart lysate. Could you please provide this information for ab109908 Complex II Enzyme Activity Microplate Assay Kit? I look forward to your reply. Please do not hesitate to contact us if you need any more advice or information. Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Abcam Scientific Support

    Answered on Nov 29 2012

    Question

    Yes, that is the correct address. It's usually best to add a few lines to help it get to our lab. This is the complete address:

    Read More

    Abcam community

    Verified customer

    Asked on Sep 21 2012

    Answer

    Thank you for confirming that information.

    I have processed the free of charge replacement and you should have it on Monday, hopefully by early afternoon. Your new order number is **.

    If you have any issues with the new kit, or if there is anything else I can help you with, then please let me know.

    Hope you have great weekend.

    Read More

    Abcam Scientific Support

    Answered on Sep 21 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2023 Abcam plc. All rights reserved.