Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909)

Overview

  • Product nameComplex IV Human Enzyme Activity Microplate Assay Kit
    See all Complex IV kits
  • Detection methodColorimetric
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts, Tissue
  • Assay typeEnzyme activity
  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Cow, Human, Pig
    Does not react with: Mouse, Rat
  • Product overview

    The Complex IV Human Enzyme Activity Microplate Assay Kit is used to determine the activity of cytochrome c oxidase in a human sample with greater 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.

  • Notes

    Store Tube 1, detergent, and microplate at 4°C – DO NOT FREEZE. Store Reagent C at -20°C or at -80°C for long-term storage.

  • Tested applicationsSuitable for: Functional Studiesmore details
  • PlatformMicroplate

Properties

Applications

Our Abpromise guarantee covers the use of ab109909 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.

Complex IV Human Enzyme Activity Microplate Assay Kit images

  • 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.
  • 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.

Protocols

References for Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909)

This product has been referenced in:
  • Chen W  et al. RIP1 maintains DNA integrity and cell proliferation by regulating PGC-1a-mediated mitochondrial oxidative phosphorylation and glycolysis. Cell Death Differ N/A:N/A (2014). Read more (PubMed: 24583643) »
  • Ahuja P  et al. Divergent mitochondrial biogenesis responses in human cardiomyopathy. Circulation 127:1957-67 (2013). Read more (PubMed: 23589024) »

See all 7 Publications for this product

Product Wall

Thank you for contacting us. The lab informed me that sample extracts can be frozen at -80oC when undiluted. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

complex IV enzyme activity assay kit

Good Average 3/5 (Ease of Use)
Abreviews
this kit was disigned for use of human samples. I was try to use it to test the porcine kidney and myocardio. The good news is it worked for both tissues.
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Submitted Mar 18 2014

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 who...

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Thank you for your patience in awaiting our reply. This kit was developed using tissue or cell celtures, and we would expect a higher reading than you got in those types of samples.

Technically no reason why you couldn’t use blood &ndas...

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Thank you for contacting us.


Technically no reason why you couldn’t – however I think the concentration of extracellular COX would be very low in serum or plasma even in cases of tissue damage.

If you are anticipat...

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Thank you for contacting us.
We following kits available for complex III. DO you think this the one you are looking for;
http://www.abcam.com/mitotox™-complex-iii-oxphos-activity-microplate-assay-ab109905.html
I hope this information is he...

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Thank you very much for having patience!

I looked at the data -the curve lost linearity after 50 sec because the substrate presumably is starting to deplete (there is less reduced cytochrome c as the enzyme clearly is using it). A way around t...

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Thank you very much for your email.

I can confirm that I am still working on your inquiry. I will send you the solution soon.

Thank you very much for having patience!

Thank you for contacting us.
In order to help you with your question we need more information about the protocol used. Please submit any raw data you have.

We will look forward to hearing from you soon.

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.

Extinc...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"