Overview

  • Product name

    Complex III Immunocapture Kit
    See all Complex III kits
  • Assay type

    Quantitative
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    250 µg, 500 µg or 750 µg monoclonal antibodies irreversibly crosslinked to protein G-agarose beads which can immunocapture ~25 µg, ~50 µg or ~75 µg respectively of Complex III from heart mitochondria.


     


    ab109800 Complex III Immunocapture Kit allows isolation of the ubiquinol-cytochrome c oxidoreductase complex from small amounts of tissue. This facilitates subsequent analysis of assembly state, activity and the extent of post translational modifications including oxidative damage that occur with aging. Uses for ab109800 include research on genetic mitochondrial disease, where complex III may have an additional role in the stabilization of complex I.


     


    Note: The immunocapture protocol for this kit requires Abcam detergent lauryl maltoside (ab109857/MS910).


     


     

  • Notes

    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.

Properties

Images

  • Complex III immunoprecipitation using antibody ab109862/MS301 crosslinked to protein G-agarose beads as product ab109800.

Protocols

References

This product has been referenced in:

  • Desmurs M  et al. C11orf83, a mitochondrial cardiolipin-binding protein involved in bc1 complex assembly and supercomplex stabilization. Mol Cell Biol 35:1139-56 (2015). Read more (PubMed: 25605331) »
  • Korde AS  et al. Primary role of mitochondrial Rieske iron-sulfur protein in hypoxic ROS production in pulmonary artery myocytes. Free Radic Biol Med 50:945-52 (2011). Read more (PubMed: 21238580) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.

Yes, the antibody coated beads can pull down the whole complex but need to make sure your customer needs to use Lauryl-β-D-maltoside (https://www.abcam.com/10-Lauryl-Maltoside-Solution-ab109857.html) to solubilized his samples. other detergents may rupture the complex structure.

This antibody is a capture antibody, we do not know the exact targeted subunit.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

The 250 ug size kit contains 25 uL of beads in a total volume of 400 uL. The buffer is 1X Hepes-buffered saline with 0.02% sodium azide preservative.

I hope this is helpful. Please contact us again if you have any further questions.

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Answer

We put 25 uL of beads in each unit.

I hope this is helpful. Please contact me again if you have any further questions.

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Question
Answer

Thank you very much for your call yesterday and for your patience while I have been in touch with the lab regarding your enquiry.

After the IP, you can use Ab110252 (UQCRC1), Ab14745(UQCRC2), Ab110255 (UQRCRQ) or Ab14746(UQCRFS1) as follow up antibodies in Western blot.

I hope that this information will be useful, but if you have any further questions or need anything else, please let me know and I'll be happy to help.

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Answer

The MitoSciences laboratory recommends the following: Use 1 mg of brain tissue lysate, eluting into the minimum volume possible. A glycine buffer elution procedure allows the reuse of the beads. If you wish to analyze the product by Western blot, we can scale back the amount of material required. So using whole tissue I think you can be very confident using 1 mg of tissue or less. However, be aware of possible mouse capture antibody elution from the beads into the sample when blotting. Ways to resolve this problem: (1) Do not include reducing agent in sample during electrophoresis. This should maintain eluted antibody at 200 kDa during electrophoresis. (2) Western blotting – Blot with rabbit antibodies or use a secondary detection system that recognizes only the native mouse antibody being used to Western blot.

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Answer

Thank you for contacting us. The protocol recommends 300 ug minimum and 5 mg ideally, but these figures refer to isolated mitochondria, and not intact tissue. I have asked the laboratory to send this information, the minimum amount of tissue required, and I will forward this when I receive it. If you are interested in isolating mitochondria, which will make the immunocapture more efficient and the yield more pure, I recommend considering the mitochondria isolation kits, ab110168 and ab110169.

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