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/lateral-flow/complex-iv-human-enzyme-activity-dipstick-assay-kit-ab109876.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 Human Enzyme Activity Dipstick Assay Kit (ab109876)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (11)References (13)

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 Human Enzyme Activity Dipstick Assay Kit (ab109876)
  • Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
  • Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876

You may also be interested in

Assay
Product image
Complex I Enzyme Activity Dipstick Assay Kit (ab109720)
Assay
Product image
Complex II Immunocapture Kit (ab109799)
Lysate
Mouse brain tissue lysate - mitochondrial extract (ab110351)

View more associated products

Overview

  • Product name

    Complex IV Human Enzyme Activity Dipstick Assay Kit
    See all Complex IV kits
  • Sample type

    Cell culture extracts, Tissue
  • Assay type

    Sandwich (quantitative)
  • Species reactivity

    Reacts with: Cow, Human
    Does not react with: Mouse, Rat
  • Product overview

    Contains 30 or 90 dipsticks and necessary components to quantify the activity of the cytochrome c oxidase enzyme complex from human and bovine samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.



    The isolation of mitochondria is not necessary for the performance of this assay. In this kit the specificity of anti-COX monoclonal antibodies (mAbs) is combined with traditional methods for determining COX enzyme activity by histochemical methods and in-gel activity assays. First, the COX enzyme complex is immnocaptured (i.e immunoprecipated in active form) on the dipstick. Second, the dipstick is immersed in COX activity buffer containing reduced cytochrome c and di-amino benzidinetetrachloride (DAB), which serves as the reporter of COX activity. Immunocaptured COX oxidizes cytochrome c, which then oxidizes DAB to form a red-colored precipitate at the COX antibody line on the dipstick. In addition to being quick, the reaction is cyanide-sensitive. The signal intensity of this precipitate corresponds to the level of COX activity in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.

  • Notes

    Store dipsticks at room temperature in their provided container and out of direct sunlight. High humidity conditions should be avoided.

    Store Buffer A, B, and C at 4°C or at -20°C for long term storage.

    Store Tubes 1 and 2 at -80°C; they can also be aliquoted upon receipt to prevent freeze/thaw cycles.

    Tube 3 can be stored at room temperature.

     

    Range of complex IV / cytochrome c oxidase assay kits

    Biochemical assay - ab239711

    Immunocapture with biochemical assay (plate-based) - ab109911 (rodent)  and ab109909 (human)

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

    Immunocapture with biochemical assay and ELISA - ab109910 (human) 

    ELISA - ab179880 (human)

  • Tested applications

    Suitable for: Functional Studiesmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 30 tests
    96-well microplate 2 units
    Buffer B (10X Blocking solution) 2 x 1ml
    Dipsticks 1 x 30 units
    Extraction Buffer (ab260490) 1 x 15ml
    Tube 1 (DAB 100X stock) 1 x 100µl
    Tube 2 (reduced cytochrome c 20X stock) 1 x 500µl
    Tube 3 (Buffer for activity assay) 1 x 10ml
    Wash buffer 1 x 1.5ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Cytochromes
  • Alternative names

    • cco
    • COX
    • Cytochrome c oxidase
    • Cytochrome oxidase
  • Database links

    • Entrez Gene: 4512 Human
    • Entrez Gene: 4513 Human
    • Entrez Gene: 4514 Human
    • Entrez Gene: 1345 Human
    • Entrez Gene: 1351 Human
    • Entrez Gene: 1329 Human
    • Entrez Gene: 1349 Human
    • Entrez Gene: 1337 Human
    • Entrez Gene: 1327 Human
    • Entrez Gene: 1340 Human
    • Entrez Gene: 1350 Human
    • Entrez Gene: 9377 Human
    • Entrez Gene: 1346 Human
    • SwissProt: P00395 Human
    • SwissProt: P00403 Human
    • SwissProt: P00414 Human
    • SwissProt: P13073 Human
    • SwissProt: P12074 Human
    • SwissProt: P15954 Human
    • SwissProt: P24311 Human
    • SwissProt: P10176 Human
    • SwissProt: P14854 Human
    • SwissProt: P09669 Human
    • SwissProt: P24310 Human
    • SwissProt: P20674 Human
    • SwissProt: P10606 Human
    see all

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109876 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies
Use at an assay dependent dilution.
Notes
Functional Studies
Use at an assay dependent dilution.

Images

  • Functional Studies - Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
    Functional Studies - Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)

    Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. 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.

  • Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
    Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)

    Figure 1. An example using ab109876 to measure Complex IV activity in fibroblast protein extracts. Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 100 µg of fibroblast protein extract.

  • Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876
    Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876

    Figure 2. An example using ab109876 to measure Complex IV activity in fibroblast protein extracts. Based on the standard curve, 50 µg of protein extract were loaded onto a dipstick for each sample. The figure shows four developed dipsticks, a control sample (1) and four unknowns (2-6). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 15-61% of normal Complex IV activity levels.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (13)

Publishing research using ab109876? Please let us know so that we can cite the reference in this datasheet.

ab109876 has been referenced in 13 publications.

  • Durhuus JA  et al. Simvastatin improves mitochondrial respiration in peripheral blood cells. Sci Rep 10:17012 (2020). PubMed: 33046789
  • Zhao T  et al. PISD is a mitochondrial disease gene causing skeletal dysplasia, cataracts, and white matter changes. Life Sci Alliance 2:N/A (2019). PubMed: 30858161
  • Nimmo GAM  et al. Bi-allelic mutations of LONP1 encoding the mitochondrial LonP1 protease cause pyruvate dehydrogenase deficiency and profound neurodegeneration with progressive cerebellar atrophy. Hum Mol Genet 28:290-306 (2019). PubMed: 30304514
  • Li H  et al. Mitochondrial dysfunction and mitophagy defect triggered by heterozygous GBA mutations. Autophagy 15:113-130 (2019). PubMed: 30160596
  • Rodríguez-García ME  et al. An innovative strategy to clone positive modifier genes of defects caused by mtDNA mutations: MRPS18C as suppressor gene of m.3946G>A mutation in MT-ND1 gene. Hum Genet 136:885-896 (2017). PubMed: 28526948
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 11 Abreviews or Q&A

Question

Inquiry: For the protein quantitation of samples in buffer A, which assays besides BCA are compatible with buffer A. will Bradford assay work?

Read More

Abcam community

Verified customer

Asked on May 17 2013

Answer

Thank you for your inquiry.

I am sorry to confirm that we are not sure what other protein assays will be compatible.

I think Bradford Assay is sensitive to detergents in general and therefore most probably not suitable.

If you like this could be quickly tested by using the extraction buffer alone in the Bradford reagent to see if it changes color.

Another option would be to measure with UV 280 for protein samples. This will also work without any additional material or kits.

I am sorry that I did not have a straight forward answer for you on this occasion and hope this information is nevertheless helpful.

Read More

Abcam Scientific Support

Answered on May 17 2013

Question

can i purchase cytochrome c sepatately?

Read More

Abcam community

Verified customer

Asked on Apr 18 2013

Answer

Thank you for contacting us.

Cytochrome c is available in our catalog, ab140219

Read More

Abcam Scientific Support

Answered on Apr 18 2013

Question

Hi, I'm looking to assay three cell types for mitochondrial protein quantity and activity. I would like to use the dipstick assays by MitoSciences (ab109720, ab109876, ab109722, and ab109877) but really do not need 30 tests of each--only 3. Is there an "assorted" pack I could purchase that just has one (or 5, or 10) of each of the four assays that are offered?

Read More

Abcam community

Verified customer

Asked on Aug 02 2012

Answer

Thank you for contacting us.

We don't have exactly what you're looking for unfortunately, but we do have MetaPath™ Mito Disease 4-Plex Dipstick Array, ab109879.

The ab109879 is a novel array that allows for the simultaneous quantification of 4 key enzymes whose expression is down-regulated in different mitochondrial diseases: Complex I, Complex IV, PDH and Frataxin. The array is comprised of 4 monoclonal capture antibodies striped onto a dipstick, and the immunocaptured sample is detected using 4 monoclonal detector antibodies that recognize different epitopes on the target enzymes. This array is very rapid (total assay time including sample prep is less than 1 hour) and very sensitive, and as with all Abcam's assays, isolation of mitochondria is not required.

This array is suitable for testing a variety of sample types. Not only is it useful for measuring samples that are deficient in the 4 enzymes due to genetic mutations, it is also useful for measuring changes in the expression of the enzymes due to drug treatments or other manipulations.

https://www.abcam.com/MetaPath™-Mito-Disease-4-Plex-Dipstick-Array-ab109879.html

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 02 2012

Question

Hi,
Thank you very much!!

Now I just need to convince my money-keeper that we need to do these experiments ;o)

Best regards

Read More

Abcam community

Verified customer

Asked on Jun 06 2012

Answer

I'm glad you have found the information helpful. If you do have any further questions please do let me know.

Until then I wish you all the best and good luck with the money-keeper :)

Read More

Abcam Scientific Support

Answered on Jun 06 2012

Question

Dear Abcam.

Thank you very much for your reply it was very usefull!!

I am also wondering whether I can use frozen human cells on your'Complex IV human enzyme activity microplate assay kit' (ab109909) or'Complex IV human specific activity microplate assay kit' (ab109910)?

Best regards

Read More

Abcam community

Verified customer

Asked on Jun 01 2012

Answer

I now have further information to share with you in regards to the kits Complex IV human enzyme activity microplate assay kit (ab109909)andComplex IV human specific activity microplate assay kit (ab109910).

As with the Complex IV human enzyme dipstick (ab109876) which you were interested in, these kits are also compatible with the sort of sample you are hoping to use. You could make a pellet of cells and freeze them at -80C as a dry pellet. There is in fact no need to freeze the cells viable.

What we recommend is to do the activity assay on the day the cells are treated with detergent, otherwise they may run the risk of degradation of the sample while stored in the provided detergent at -80C.

I hope this information has been of help. If you have any further information please do not hesitate to contact us again.

Read More

Abcam Scientific Support

Answered on Jun 01 2012

Question

Dear Abcam.

I am very interested in your dipstick activity assay kit. Especially the complex IV human enzyme dipstick. However, I have a question about this.

Do you have any experience with the accuracy of the complex IV activity in samples which have been frozen?

Specifically I have human periferal blood mononuclear cells dissolved in 70% fetal calf serum, 25% RPMI and 5% DMSO frozen at -80*C and -120*C.

Do you have experience with activity accuracy of any of your other dipstick assay kits on frozen samples?

Best regards

Read More

Abcam community

Verified customer

Asked on May 28 2012

Answer

Thank you for contacting us and your interest in our products.

The Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)was validated by MitoScienceswith frozen cell pellets (non viable) from patients with SIRF1 and COX10 mutations leading to complex IV deficiency. The attached paper shows the validation data. Your samples should work well with the kitsince the PBMCs were frozen viable.In order to use them with the kit you would first need tothaw them, wash well the FBS with PBS by centrifugation, then solubilize the pellet according to the protocol instructions and measure protein concentration either by 280nm (UV) or by BCA assay before loading on the dipstick.

The paper attached also shows accuracy with the complex I activity dipstick.

I hope this information has been of help. If you have any further questions please do not hesitate to contact us again.

Read More

Abcam Scientific Support

Answered on May 28 2012

Question

Hi,

I'm analysing cultured fibroblasts by western blotting (protein extracted in RIPA buffer) and complex I/IV dipstick assays (in buffer A from ab109876 or ab109720). I'm wondering if it's possible to use the buffer A samples for western blotting? if yes, any additional steps required before loading or it's the same like how i treat my RIPA samples?

Thanks

Regards,

Read More

Abcam community

Verified customer

Asked on May 21 2012

Answer

Thank you for contacting us.
Yes these samples can be used for Western blotting just as one would with RIPA, the extraction detergent in A is very similar to RIPA (NP40 plus others) but it is more native.
The only thing that I would say is that when used concentrated and diluted in the Western blot sample buffer, detergent may affect SDSPAGE migration at the very bottom of the lane (much less than 10kDa) but all other migration should be normal.
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 May 21 2012

Question

buffer B missing
kit for 90 tests

Read More

Abcam community

Verified customer

Asked on Mar 29 2012

Answer

Thank you for your phone call.

I am sorry this product did not contain buffer Bas 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.

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 Mar 29 2012

Question

I recently purchased a Complex IV activity dipstick assay from your company and had a few questions about data analysis.

The samples used were of whole blood and they were prepared according to the protocol abcam suggested: Making a 1:3 dilution with the extraction buffer, allowing it to sit on ice for 20 min and then spinning it in a microcentrifuge at 13,000-16,000rpm for 20min. The supernatant was removed and 0.3-9ul of the supernatant was added per dipstick

The assay protocol for the Complex IV activity assay was followed as indicated by the kit insert and a dipstick reader was used to analyze each stick.

Since the reader is new, the stick was read by placing the nitrocellulose membrane inside the machine with the wicking pad facing the door.

However, The data I have obtained is confusing to interpret. I am following each mABS of each peak, which I am expecting only two, one for the positive control and another for the activity complex IV. I assume that the complex IV dipstick has a positive control.

I have attached some data I have obtained on whole blood samples using a range of concentrations. The results are puzzling. I am not sure which peak belongs to Complex IV and which peak belongs to the positive control. Is there an approximate range I should expect Complex IV to show up? I understand that perhaps wicking of a sample can be uneven and therefore give variable results but there must be some range I should be expecting the peak.

Read More

Abcam community

Verified customer

Asked on Feb 29 2012

Answer

Thank you for contacting Abcam

I sent you query to our lab to ask their opinion on your data and I have attached the file edited with whatthey consider are the positive peaks (without looking at the actual dipsticks). The positive peaks are highlighted in green. The signal is very weak, but the ones highlighted in green are the only ones that show increase in activity with higher loading. The dipstick reader will allocate number of peaks. The peak of interest will not always appear in the same number. Whatyou should look for is the peak at a particular position of X. In this case, they need to look at the peak between 460 and 490 (which in some dipsticks is peak 1, in others is peak 2 and in others is peak 3).

When loading blood, we typically don’t load it by the protein concentration, since most of it is going to be hemoglobin present in red blood cells and albumin present in the serum which are not representative of the mitochondria content in blood (present in PBMCs and platelets).You should load by uL of extract (and not by ug). In the graph below, we have the data you should expect by loading 90uL of blood extract, 45uL……in a 1:2 dilution series. The data was obtained with pooled blood from 3 healthy volunteers. Up to 100uL of volume could be loaded on the dipstick. The 90uL loading was achieved by combining 90uL of extract with 10uL of 10x blocking.

For optimal results, it is also important for you to use fresh blood (not frozen) or if this is not possible, blood should at the very least be stored at -80C viable by adding 10% DMSO just like you would on cultured cells). We have only tested the dipsticks with fresh blood and so cannot guarantee their performance on frozen viable blood, but theoretically it should work well. Also for higher quality data it is recommended to isolate PBMCs from the blood and use that in the assay as opposed to whole blood samples.

Read More

Abcam Scientific Support

Answered on Feb 29 2012

Question

The data we get from reading abcam dipsticks used on lysed whole blood has 5-6 peaks. We would like to know what each peak refers to and which to consider when analyzing data.   Is it necessary to change any settings on the reader when reading dipsticks from different assay kits?   Can you provide us with an example of analyzed data obtained from the dipstick reader?   The dipstick kits we are having trouble with are… ab109720 – Complex I Enzyme Activity Dipstick Assay Kit ab109876 – Complex IV Enzyme Activity Dipstick Assay Kit   Thank you,

Read More

Abcam community

Verified customer

Asked on Feb 17 2012

Answer

Thank you for contacting Abcam regarding this issue.

On occasion the reader will see more profiles which cannot be seeing with the naked eye on the dipstick. This is due sometimes to imperfections of the nitrocellulose or to uneven wicking. For activity assays the antibody line is stripped at about 7mm from the nitrocellulose edge of the dipstick (or lower end of the dipstick). In new readers, the dipstick is inserted with the wicking pad towards the door of the instrument and the nitrocellulose towards the inside of the instrument. The reader will read from the inside of the instrument (lower section of the dipstick) to the outside of the instrument (upper section of the dipstick). Therefore the lower section of the dipstick will appear towards the left of the absorption profile. The reader will pick up all peaks in a sample .



We strongly recommend creating a standard curve using 7 to 8 dipsticks for covering the working range. In this way you will be able to know which peaks are background and which are signal. Following the protein concentration ranges as defined in Table 1 of the protocol, generate a standard curve using a positive control sample. We recommend performing a 1:2 serial dilution with 1 part Buffer A to 1 part Buffer B in a total volume of 50 µl.



Complex IV dipsticks may sometimes develop a percipitate about 2mm from the edge, this should be ignored.



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 17 2012

1-10 of 11 Abreviews or Q&A

  •  Previous
  • 1
  • 2
  • 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.