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

Complex IV Human Specific Activity Microplate Assay Kit (ab109910)

Submit a review Q&A (8)References (11)
Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
  • Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
  • Sandwich ELISA - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: Cell culture extracts, Tissue

Overview

  • Product name

    Complex IV Human Specific Activity Microplate Assay Kit
    See all Complex IV kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture extracts, Tissue

  • Assay type

    Enzyme activity

  • Species reactivity

    Reacts with: Human

  • Product overview

    This rapid multiplexing microplate kit is used to determine both the activity and quantity of cytochrome c oxidase in a human sample with greater speed and simplicity. The ratio of the two is a measure of specific activity. The COX enzyme is immunocaptured within the wells of the microplate and the activity is determined by following the oxidation of reduced Cytochrome c colorimetrically by the absorbance change at 550 nm. Subsequently, in these same wells the quantity of the enzyme can then be determined by adding a COX specific detector antibody and the appropriate alkaline phophatase conjugated antibody; this phosphatase changes a substrate from colorless to yellow at 405 nm. This reaction takes place in a time dependent manner proportional to the amount of protein captured in the wells. Included in this kit for performance of the multiplexing assay are buffers, detergent, substrates, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.

  • Notes

    Range of complex IV / cytochrome c oxidase assay kits

    Biochemical assay - ab239711

    Immunocapture with biochemical assay (plate-based)*** - ab109911 (rodent)  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) (this kit)

    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

Images

  • Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
    Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
    Functional study using Complex IV Human Specific Activity Microplate Assay Kit (ab109910).
  • Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
    Functional Studies - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
    Functional study using Complex IV Human Specific Activity Microplate Assay Kit (ab109910).
  • Sandwich ELISA - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)
    Sandwich ELISA - Complex IV Human Specific Activity Microplate Assay Kit (ab109910)

    Principle of Sandwich ELISA.

References (11)

Publishing research using ab109910? Please let us know so that we can cite the reference in this datasheet.

ab109910 has been referenced in 11 publications.

  • Perez JM  et al. Proteomic Analysis Reveals Low-Dose PARP Inhibitor-Induced Differential Protein Expression in BRCA1-Mutated High-Grade Serous Ovarian Cancer Cells. J Am Soc Mass Spectrom 33:242-250 (2022). PubMed: 34958553
  • Yao PJ  et al. Mitochondrial Electron Transport Chain Protein Abnormalities Detected in Plasma Extracellular Vesicles in Alzheimer's Disease. Biomedicines 9:N/A (2021). PubMed: 34829816
  • Carling PJ  et al. Deep phenotyping of peripheral tissue facilitates mechanistic disease stratification in sporadic Parkinson's disease. Prog Neurobiol 187:101772 (2020). PubMed: 32058042
  • Richter F  et al. ROMO1 is a constituent of the human presequence translocase required for YME1L protease import. J Cell Biol 218:598-614 (2019). PubMed: 30598479
  • Aich A  et al. COX16 promotes COX2 metallation and assembly during respiratory complex IV biogenesis. Elife 7:N/A (2018). PubMed: 29381136
View all Publications for this product

Customer reviews and Q&As

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

1-8 of 8 Abreviews or Q&A

Question

step 10.2.8 ....should we leave the 200ul of assay solution in the plate and cover overnight at 4C or empty it out, or add wash solution?

Read More
Answer

Please leave the 200uL of assay solution in the wells, cover and incubate at 4C overnight.

Read More

Question

I have questions regarding the Complex IV Human Specific Activity Microplate Assay Kit (ab109910) and the NADH Dehydrogenase (Complex I) Human Profiling ELISA kit (ab124535). Can the detergent for ab109910 be used in place of the extraction buffer for ab124535 and vice versa? Also, would it be possible to place an order for only the detergent or extraction buffer without purchasing an entire kit?

Read More
Answer

Yes, we use 10% detergent in ab109910 which is available as ab109857 so the final is 1%. This detergent is pretty mild and yields native functional enzymes.

Also, a final of 1.5% could be used (i.e. extraction buffer) without significantly changing the results or activity yield.

Read More

Question

“I would like to assay complex 4 activity in primary neurons after different treatments. On the first time I have used that kit I got some samples that show a first slope (increasing wthin the first minute) then a second slope decreasing fast and a third slope that is deceasing and maintained until the end.

Which slope should I take into account? the only slope that appears in all the samples is the last. Why do I get the first slope that increases?

Read More
Answer

The slope to measure is the fast decreasing slope. The cytochrome c oxidase activity assay measures the capacity of the immunocapture enzyme for oxidizing reduced cytochrome c. Reduced cytochrome c is measured at 550nm. A reduction in the reading means that the reduced cytochrome c is becoming oxidized.

Read More

Question

Customer contacted us if it was ok to perform step 7. A) incubation O/N at 4oC instead of 3 hours at RT.

Read More
Answer

We have not tested the enzyme activity portion of the assay (ab109910) after overnight incubation at 4C. My suggestion is to not do it and stick to the conditions set out in the protocol = 3 hours incubation of sample at RT – then perform the activity assay portion (this should be done on the same day the sample was extracted) – then leave overnight at 4C (with the activity buffer in the wells) and do the Sandwich ELISA portion of the assay the next day. With this format we know the assay performs optimally.

Read More

Question

i have some questions about Complex IV Human Specific Activity Microplate Assay Kit (ab109910);

First, i'm doing cell culture with C2C12 cell line (mouse), i want to measure cytochrome C activity; i use 4 X10 5 cells /well in 12 well- PLATES, than i extract my proteins with RIPA buffer, my questions are :

1. Is there any kit to measure cytochrome C activity special mouse not human §

2. If yes , can i use this kit with my experimental conditions written above ( i mean with C2C12 cell line , the number of cell 4 X 10 power 5 cells )

3. This kit ; i must use it when i collect my cells with PBS ( cells will be in suspension ) or after i will extract my proteins using RIPA buffer ?

Note : i hade already recommanded mitochondria extraction kit from your compagny but i can't use it to extract mitochondria because my number of cells is so low and i can't increase it, so can i use a kit to measure cytochrome C activity with these conditions ;

Read More
Answer

Thank you for contacting us.

You may be interested in our cytochrome c protein quantity microplate assay kit, ab110172.

https://www.abcam.com/ps/products/110/ab110172/documents/ab110172%20Cytochrome%20c%20Protein%20Quantity%20microplate%20assay%20kit%20(Website).pdf

It will work on cultured cell extracts from 0.5-50 ug/ml (3 x 10^4 - 3 x 10^5 cells/ml). You can also use the supplied detergent to solubilize samples.

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

Question

I have a few questions about this kit:
1. What is the product or kit recommended to produce the pure mitochondrial preparation for use as a sample in this kit?
2. Does 'Solution 1' contain succinate for use as electron donor?
3. Can BCA be used to determine protein concentration of samples for use in this kit?

Read More
Answer

Answer 1. Mitochondrial isolation can be performed according to kits ab110169 or ab110171.

2. Answer 2. Solution 1 does not contain succinate; instead the cytochrome c has been reduced with DTT and then purified.

Answer 3. BCA can be used to determine protein concentration.

Read More

Question

I am interested in buying the kit:
https://www.abcam.com/Complex-IV-Human-Specific-Activity-Microplate-Assay-Kit-ab109910.html
I want to do the assay with human primary neurones and astrocytes.
- How many cells do I need to have to get appropriate results or how much protein?
- How do I need to lysate them? I need to do BSA protein assay previously to the complex IV assay if I extract my cells with the solution 1 would that be compatible with the BSA assay?
-What should I use as a positive control sample?

Thanks a lot for your help,

Kind regards,

Read More
Answer

Thank you for contacting us.

Please find attached the sample preparation guide recommended by our Mitoscience colleagues.

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

Question

Could you provide the MSDS for ab109910 and ab110671.

Read More
Answer

Thank you for calling Abcam. Please find attached the MSDS for ab109910 and ab110671. If there is anything else I can do, please let me know.

Read More

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