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Pyruvate dehydrogenase (PDH) Profiling ELISA Kit (ab110174)

Submit a review Q&A (4)References (2)
ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)
  • Sandwich ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)

Key features and details

  • Sample type: Cell culture extracts, Tissue
  • Detection method: Colorimetric
  • Assay type: Sandwich (qualitative)
  • Reacts with: Mouse, Rat, Cow, Human

Overview

  • Product name

    Pyruvate dehydrogenase (PDH) Profiling ELISA Kit
    See all Pyruvate dehydrogenase kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture extracts, Tissue

  • Assay type

    Sandwich (qualitative)

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human

  • Product overview

    Abcam’s Pyruvate dehydrogenase (PDH) in vitro ProfilingELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the measurement of Pyruvate dehydrogenase (PDH) in Human, bovine, mouse, and rat whole tissue or cell lysate samples.

    Capture antibodies are pre-coated in the wells of modular microplates, which can be broken into 8-well strips. This assay is a “sandwich” ELISA, where the PDH enzyme is purified and immobilized by an anti-PDH capture antibody pre-coated in the microplate wells. The amount of captured PDH is determined by adding a second (detector) anti-PDH antibody which binds to the captured PDH hat a different epitope. This is followed by binding of an HRP conjugated goat anti-mouse antibody that binds the detector anti-PDH antibody. The detector-bound HRP then changes the colorless HRP development solution to blue and the color intensity (absorbance) is proportional to the amount of PDH captured. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state.

  • Notes

    5X Stabilizer should be stored at -20°C. Remainder of kit should be stored at 4°C. When stored as recommended the kit is stable for 6 months.

     

    PDH is the key regulatory enzyme of cellular metabolism because it links the TCA cycle and subsequent oxidative phosphorylation with glycolysis and gluconeogenesis as well as with both lipid and amino acid metabolism. PDH activity is regulated primarily by PDK-dependent phosphorylation and PDP-dependent dephosphorylation of PDH. Phosphorylation inactivates PDH whereas dephosphorylation activates PDH. Phosphorylation occurs at Serines 232, 293, and 300 of the human E1α subunits.

  • Platform

    Microplate

Properties

Images

  • ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)
    ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)
    An example of the quantity of PDH capture from a HepG2 cultured cell lysate. The sample was diluted to show that over this range of concentrations that can be used. Each sample was measured in 6 replicates. Bars show standard deviations.
  • Sandwich ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)
    Sandwich ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Microplate Assay Kit (ab110174)
    Abcams' protein quantity microplate assays use the well-established sandwich ELISA format, whereby capture and detector antibodies are used to immobilize and then quantify a target protein or enzyme. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state. 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, a second monoclonal antibody, against a different epitope on the target, is added to the well. This detector antibody is either directly labeled with biotin, or a biotin-labeled goat anti-mouse secondary is added. Substrate plus HRP or AP conjugated to streptavidin provide a colorimetric signal that is readable by any plate readers capable of standard ELISA absorbance measurements.

References (2)

Publishing research using ab110174? Please let us know so that we can cite the reference in this datasheet.

ab110174 has been referenced in 2 publications.

  • Zager RA  et al. Renal Cortical Pyruvate Depletion during AKI. J Am Soc Nephrol N/A:N/A (2014). ELISA ; Mouse . PubMed: 24385590
  • Yoshioka J  et al. Deletion of thioredoxin-interacting protein in mice impairs mitochondrial function but protects the myocardium from ischemia-reperfusion injury. J Clin Invest 122:267-79 (2012). PubMed: 22201682

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Question

1) To which subunit does the antibody bind?

2) Does the antibody recognize a linear epitope or a conformational epitope?

3) Does the antibody bind to the active site on enzyme?

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Answer

To answer your previous questions:

1) To which subunit does the antibody bind?
E2/E3bp

2) Does the antibody recognize a linear epitope or a conformational epitope?
Conformational epitope

3) Does the antibody bind to the active site on enzyme?
The enzyme continues to be active after immunocapture with the antibody.

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Question

1. Under 'Preparation of Samples', Step 4, can the supernatant be stored at -80 degrees C at this point?
2. Under 'Assay Method', Step 2, can the plate be incubated overnight at 4 degrees C instead of at room temp for 3 hrs?

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Answer

The answer to both questions is YES.

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Question

Needs to quantify the PDH in samples, the only PDH standard available is from pig. Can the antibodies in the kit detect porcine PDH?

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Answer

Thank you very much for your call last week and for your patience while I have been in touch with the lab about this kit.

The antibodies used in the kit were actually raised against the pig protein,so we expect it to work with the pig protein. Wehave alsoreceived reports from usersstating thatthe kit works with pig, though we have not tested the kit with pig samples in our own lab.

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

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Question

I need a kit which will measure preferably enzymatic activity but also could measure protein quantity. My question is, will the 8M urea, 4% CHAPS lysis buffer that my samples are in affect these products?

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Answer

Thank you for contacting ABcam in regards to the lysis buffer that your tissues are in and how that will work with our PDH kit.

These kits will not function well with samples in such a harsh buffer. It is recommended that samples are in PSB buffers. You may wish to use a buffer exchange system such as a dialysis tubing to remove the 8M urea and replace it with PBS. Thereby enabling you to use these samples in these kits.

I hope that you have found this information useful. Please let me know if you have any questions.

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