Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)

Overview

  • Product nameComplex IV Human Enzyme Activity Dipstick Assay Kit
    See all Complex IV kits
  • Sample type
    Cell culture extracts, Tissue
  • Assay typeSandwich (quantitative)
  • Species reactivity
    Reacts with: Cow, Human
  • 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.

  • Tested applicationsSuitable for: Functional Studiesmore details
  • PlatformReagents

Properties

Applications

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.

Complex IV Human Enzyme Activity Dipstick 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. 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.
  • 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.
  • 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

References for Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)

This product has been referenced in:
  • Calvo SE  et al. Molecular diagnosis of infantile mitochondrial disease with targeted next-generation sequencing. Sci Transl Med 4:118ra10 (2012). Read more (PubMed: 22277967) »
  • Fassone E  et al. FOXRED1, encoding an FAD-dependent oxidoreductase complex-I-specific molecular chaperone, is mutated in infantile-onset mitochondrial encephalopathy. Hum Mol Genet 19:4837-47 (2010). Read more (PubMed: 20858599) »

See all 7 Publications for this product

Product Wall

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.

I...

Read More

Thank you for contacting us.

Cytochrome c is available in our catalog, ab140219

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 ...

Read More

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 :)

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...

Read More

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 l...

Read More

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 th...

Read More

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.
Read More

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 h...

Read More

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. ...

Read More

1-10 of 11 Abreviews or Q&A

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