Complex IV Immunocapture Kit (ab109801)

Submit a review Q&A (3)References (9)
Immunoprecipitation - Complex IV Immunocapture Kit (ab109801)

    Key features and details

    • Assay type: Quantitative

    Overview

    • Product name

      Complex IV Immunocapture Kit
      See all Complex IV kits

    • Assay type

      Quantitative

    • Species reactivity

      Reacts with: Cow, Human

    • Product overview

      ab109801 Complex IV Immunocapture Kit allows isolation of the cytochrome c oxidase complex from small amounts of tissue.


      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 IV from heart mitochondria.


      This kit facilitates subsequent analysis of both assembly state and accrued post-translational modifications of all 13 subunits of Complex IV. The immunoprecipitated Complex IV shows cytochrome c oxidase activity while bound to the beads and this activity is fully sensitive to cyanide. Uses for ab109801 include but are not limited to examining alterations of Complex IV subunits in inherited mitochondrial diseases, Alzheimer's disease, schizophrenia and cancer.


       


      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

    • Immunoprecipitation - Complex IV Immunocapture Kit (ab109801)
      Immunoprecipitation - Complex IV Immunocapture Kit (ab109801)
      Complex IV immunoprecipitation using antibody ab109863/MS101c crosslinked to protein G-agarose beads as product ab109801

    References (9)

    Publishing research using ab109801? Please let us know so that we can cite the reference in this datasheet.

    ab109801 has been referenced in 9 publications.

    • Morscher RJ  et al. Mitochondrial translation requires folate-dependent tRNA methylation. Nature 554:128-132 (2018). PubMed: 29364879
    • Oliva CR  et al. Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells. J Biol Chem 291:24188-24199 (2016). IP . PubMed: 27679486
    • 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). PubMed: 25605331
    • van den Bosch BJ  et al. Defective NDUFA9 as a novel cause of neonatally fatal complex I disease. J Med Genet 49:10-5 (2012). PubMed: 22114105
    • Tucker EJ  et al. Mutations in MTFMT Underlie a Human Disorder of Formylation Causing Impaired Mitochondrial Translation. Cell Metab 14:428-34 (2011). PubMed: 21907147
    • Stiburek L  et al. Loss of function of Sco1 and its interaction with cytochrome c oxidase. Am J Physiol Cell Physiol 296:C1218-26 (2009). PubMed: 19295170
    • Murray J  et al. Small-scale immunopurification of cytochrome c oxidase for a high-throughput multiplexing analysis of enzyme activity and amount. Biotechnol Appl Biochem 48:167-78 (2007). PubMed: 17508937
    • Janssen GM  et al. The A3243G tRNALeu(UUR) mutation induces mitochondrial dysfunction and variable disease expression without dominant negative acting translational defects in complex IV subunits at UUR codons. Hum Mol Genet 16:2472-81 (2007). PubMed: 17656376
    • Walker UA  et al. Uridine abrogates the adverse effects of antiretroviral pyrimidine analogues on adipose cell functions. Antivir Ther 11:25-34 (2006). PubMed: 16518957

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Customer kindly contacted us to determine if this product might work with zebrafish or goldfish samples.

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    Answer

    Thank you for contacting us.

    I have been in contact with the lab regarding species reactivity of this product as you have requested. They have tested the paired antibody in zebrafish and this will definitely not work on Zebrafish or goldfish.

    While not IP or immunocapture some of the mitosciences antibodies were used in a mitochondrial zebrafish project:

    http://www.ncbi.nlm.nih.gov/pubmed?term=17761683

    Should you wish to test any Abcam products in an untested species or application we have established a testing discount program. Here is a brief description of how it works:

    The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount equal to the value of the product tested. The terms and conditions of this offer can be found here: https://www.abcam.com/AbTrial

    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

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    Question

     I would like to ask what is the minimum amount of mouse brain tissue required for Immunocapture kit Complex I (as well as complex III, and IV) so as to pull down Complex I. We would need enough material for running an SDS gel.

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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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    Question

     I would like to ask what is the minimum amount of mouse brain tissue required for Immunocapture kit Complex I (as well as complex III, and IV) so as to pull down Complex I. We would need enough material for running an SDS gel.

    Read More
    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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