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-iv-rodent-enzyme-activity-microplate-assay-kit-ab109911.pdf

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

Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (3)Q&A (34)References (82)

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.

Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
  • Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
  • Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis.

Key features and details

  • Assay type: Enzyme activity (quantitative)
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 6 hr
  • Sample type: Cell culture extracts, Tissue

You may also be interested in

Assay
Product image
Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
Assay
Product image
Complex II Enzyme Activity Microplate Assay Kit (ab109908)
Lysate
Bovine Heart Mitochondria (ab110338)

View more associated products

Overview

  • Product name

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

    Colorimetric
  • Sample type

    Cell culture extracts, Tissue
  • Assay type

    Enzyme activity (quantitative)
  • Assay time

    6h 00m
  • Species reactivity

    Reacts with: Mouse, Rat
  • Product overview

    Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911) is used to determine the activity of cytochrome c oxidase in a mouse sample with speed and simplicity. The COX enzyme is immunocaptured within the wells of the microplate and activity is determined colorimetrically by following the oxidation of reduced cytochrome c by the absorbance change at 550 nm. Included in this kit for performance of the activity assay are buffer, detergent, substrate, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.


    Complex IV assay protocol summary:
    - add samples to wells to capture enzyme and incubate for 3 hrs
    - wash wells
    - add reaction mix
    - analyze with microplate reader in kinetic mode for 120 min


    Buffer, detergent, and microplate should be stored at 4°C, Reagent C (reduced cytochrome c) should be stored at -80°C.

  • Notes

    Range of complex IV / cytochrome c oxidase assay kits

    Biochemical assay - ab239711

    Immunocapture with biochemical assay (plate-based)*** - ab109911 (rodent) (this kit) and ab109909 (human)
    *** Most popular assay format

    Immunocapture with biochemical assay (dipstick) - ab109878 (rodent) and ab109876 (human)

    Immunocapture with biochemical assay and ELISA - ab109910 (human) 

    ELISA - ab179880 (human)

     
    Other 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
    96-well microplate 1 unit
    Tube 1 (Buffer) 1 x 10ml
    Detergent 1 x 1ml
    Reagent C (Reduced Cytochrome c) 1 x 1ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Other Metabolism Assay
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Cytochromes
  • Alternative names

    • cco
    • COX
    • Cytochrome c oxidase
    • Cytochrome oxidase

Associated products

  • Positive Controls

    • 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

  • Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
    Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
    Figure 1. To determine the activity in the sample, calculate the slope by using microplate software or by manual calculations using one of the two methods shown. Compare the sample rate with the rate of the control (normal) sample and with the rate of the null (background) to get the relative Complex IV activity. (A)The rate is determined by calculating the gradient of the initial slope over the linear region. (B) The rate is determined by calculating the slope between two points within the linear region.
  • Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
    Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
    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.
  • Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis.
    Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis.Image courtesy of Yao X et al.PLoS One. 2015; 10(10): e0139416. doi: 10.1371/journal.pone.0139416.

    Mitchondiral fractions from the heart tissue of Rats infected by S. pneumoniae, or given PBS sham control, were subjected to measurements of complex I-V activities. Complex I was measured with ab109721 (top left), Complex II + III were measured using ab109905 (top right), Complex IV was measured using ab109911 (bottom left) and Complex V was measured using ab109714 (bottom right).

    Freshly isolated mitochondrial pellets were resuspened in PBS supplemented with 10% detergent provided in the kits. Protein concentrations of these mitochondrial lysates were estimated and 25 μg (for complex I, IV and V) or 100 μg (for complex II+III) mitochondrial protein was used per reaction. Enzyme activities were measured spectrophotometricly in triplicate and expressed as changes of absorbance per minute per mg protein.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (82)

Publishing research using ab109911? Please let us know so that we can cite the reference in this datasheet.

ab109911 has been referenced in 82 publications.

  • Zhu Y  et al. miR-340-5p Mediates Cardiomyocyte Oxidative Stress in Diabetes-Induced Cardiac Dysfunction by Targeting Mcl-1. Oxid Med Cell Longev 2022:3182931 (2022). PubMed: 35126811
  • 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
  • Huang L  et al. Increased fatty acid metabolism attenuates cardiac resistance to ß-adrenoceptor activation via mitochondrial reactive oxygen species: A potential mechanism of hypoglycemia-induced myocardial injury in diabetes. Redox Biol 52:102320 (2022). PubMed: 35462320
  • 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
  • Dogan AE  et al. PACT establishes a posttranscriptional brake on mitochondrial biogenesis by promoting the maturation of miR-181c. J Biol Chem 298:102050 (2022). PubMed: 35598827
View all Publications for this product

Customer reviews and Q&As

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

1-10 of 37 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 use, straight forward to standardize with cell lysates. Good specificity with microplate and results reproducible.
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 Aug 31 2018

Chronic hypoxia disrupts mitochondrial function in the guinea pig placenta

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
I used isolated protein from guinea pig placenta to perform the experiment with this kit. There is no activity detected.
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 07 2017

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

Using CHO cells. Prior to step 8, should the protein concentration be remeasured to load the recommended amounts per well, or are the recommended amounts based on the 5 mg/ml recommended concentration before adding the detergent. Also, should any of the sample preparation steps be modified for cells - it seems the protocol is tailored for tissue but on the datasheet cell culture extracts are listed as a guaranteed sample type. For these CHO cells, should I use 50 ug / 200 uL as you recommend for tissue extracts?

Read More

Abcam community

Verified customer

Asked on Aug 02 2013

Answer

The protocol makes specific reference to mitochondria and tissue extracts. When it comes to cells, there is a significant loss of protein in the pellet after extraction as you mentioned on the phone. I would recommend either re-measuring the concentration again after extraction or to load at multiple concentrations (both controls and test samples) to account for potential losses during the extraction procedure. I realize that measuring twice could be very tedious. A way around it is to resuspend the cell pellet in 8 volumes of the buffer supplied = 50uL pellet + 400uL buffer. This should be approximately 5mg/mL, then the detergent can be added which can be followed by a BCA assay.
Below is a link to a sample preparation guide for our Mitosciences ELISA assays and immunocapture activity assays, such as ab109911.
https://www.abcam.com/index.html?pageconfig=resource&rid=15500

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Aug 02 2013

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

Cytochrome C is typically isolated as a dimer. For the purpose of determining the molecular weight of the cytochrome C present after sample preparation and addition of the detergent, should I expect the cytochrome C to be dimerized and in complex with lauryl maltoside?

Read More

Abcam community

Verified customer

Asked on Jul 30 2013

Answer

We provide the cytochrome c in reduced form for this kit.
I can say the protein is active and also the detergent is mild. So if cytochrome c functions as a dimer then you can anticipate this conformation in the assay solution.
We are unsure whether the molecule will specifically be dimerized or not following addition of the detergent, but you should measure the total concentration using a standard BCA or Bradford assay and use a 5 mg/ml solution with ab109911.

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Jul 30 2013

Question

Dear All,
a customer of ours is interested in your AB10991 but in the lab they don't have a filter able to read at 550 nm. They have a filter able to read at 570 nm, do you this this kit can work also by using this filter?

Thanks in advance and kind regards.

Read More

Abcam community

Verified customer

Asked on Feb 15 2013

Answer

Thank you for your technical enquiry and interest in our products.

I have discussed this with my colleagues in the Lab. This kit (ab109911: Complex IV Rodent Enzyme) measures in a standard spectrophotometer the absorbance of reduced cytochrome c. The peak of absorbance of reduced cytochrome c is at 550 nm. This peak is unfortunately is very narrow and the readings drop quickly after 555 nm. At 570 nm the reading will be almost 0.0.

Apologies!

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Feb 15 2013

Question

I would like to order 1 unit, could you send me a product code, thanks

Regards

Read More

Abcam community

Verified customer

Asked on Dec 18 2012

Answer

Thank you for your interest.

The product code number for this item is ab140219. However, it is not published so the datasheet can't get accessed on the public site.

I would advise to place an order via e-mail: mailto:orders@abcam.com and mention that you have been discussing this special order with me.

I have ccd this response to Customer Service so they will be aware of this special order.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Dec 18 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

Hi

Would it be possible to purchase Reagent C (Cytochrome c) separately from the Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)?

Regards

Read More

Abcam community

Verified customer

Asked on Dec 17 2012

Answer

Thank you for your contact.

I have discussed your enquiry with my colleagues and can confirm that Reagent C is available for special request. The sale price is USD50.00 or GBP40.00 /unit. We currently have this item in stock.

Could you please confirm if you are still interested in getting this item and how many units you wish to purchase?

I look forward to hearing from you soon.

Read More

Abcam Scientific Support

Answered on Dec 17 2012

1-10 of 37 Abreviews or Q&A

  •  Previous
  • 1
  • 2
  • 3
  • 4
  • Next 

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.