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Overview

  • Product nameAlpha Ketoglutarate (alpha KG) Assay Kit
  • Tests
    100 x 1 assay
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • Assay typeQuantitative
  • Sensitivity
    = 0.01 nM
  • Range
    0.01 nM - 10 nM
  • Assay time
    0h 30m
  • Product overview

    Abcam's Alpha Ketoglutarate (alpha KG) Assay Kit provides a simple, sensitive and rapid means for quantifying alpha KG in a variety of samples. In the assay, alpha-ketoglutarate is transaminated with the generation of pyruvate which is utilized to convert a nearly colorless probe to both color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The alpha-Ketoglutarate Assay Kit is useful for detecting alpha-ketoglutarate in the range of 0.01 to 10 nmoles.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Alpha Ketoglutarate (alpha KG) is a key intermediate in the Krebs cycle, coming after isocitrate and before succinyl CoA. Anaplerotic reactions replenish the cycle by synthesizing alpha KG from transamination of glutamate, or through the action of glutamate dehydrogenase. Alpha KG is an important nitrogen transporter. Being a key intermediate, it is one of the organic acids measured in newborns as an indicator of inborn errors of metabolism.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceAlpha ketoglutarate (Alpha-KG) is a key intermediate in the Krebs cycle, coming after isocitrate and before succinyl CoA. Anaplerotic reactions replenish the cycle by synthesizing Alpha-KG from transamination of glutamate, or through the action of glutamate dehydrogenase. Alpha-KG is an important nitrogen transporter. Being a key intermediate, it is one of the organic acids measured in newborns as an indicator of inborn errors of metabolism.
    • Alternative names
        Alpha KG

    Applications

    Our Abpromise guarantee covers the use of ab83431 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    Functional Studies FuncS

    Alpha Ketoglutarate (alpha KG) Assay Kit images

    • Functional Studies - Alpha Ketoglutarate Assay Kit (ab83431)
      Functional Studies - Alpha Ketoglutarate Assay Kit (ab83431)
      Alpha Ketoglutarate standard curve generated using the ab83431 kit protocol.

    Protocols

    References for Alpha Ketoglutarate (alpha KG) Assay Kit (ab83431)

    This product has been referenced in:
    • Long LH & Halliwell B Artefacts in cell culture: a-Ketoglutarate can scavenge hydrogen peroxide generated by ascorbate and epigallocatechin gallate in cell culture media. Biochem Biophys Res Commun 406:20-4 (2011). Functional Studies . Read more (PubMed: 21281600) »

    See 1 Publication for this product

    Publishing research using ab83431 ? Please let us know so that we can cite the reference in this datasheet

    Product Wall

    Submit an Abreview for ab83431 Submit a Question for ab83431

    Displaying 1 - 10 of 15 results for Abreviews and Q&A

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    Votes: Highest First

    I have a doubt regarding the sample preparation for this kit.
    According with the protocol, samples should be homogenized with Ice-
    cold PBS and after that frozen with liquid nitrogen. However, I´m
    going to measure a-KG levels in cultu...

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    There are two options for the sample preparation.

    Either just rapidly homogenize cells with 100 μl of ice cold PBS or other buffer (pH 6.5-8) and then add 1-50 μl samples into duplicate wells of a 96-well plate and bring volume t...

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    The instruction book warns against pyruvate in samples causing background. I also tested the assay on pure oxaloacetate solution and saw a positive reading suggesting that this assay can react non-specifically with oxaloacetate. I believe this assay to no...

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    Thank you for contacting us. Did you compare the signal you obtained from the oxaloacetate with the results from a negative control? The control we recommend is testing without addition of the converting enzyme, in order to account for signal contributed ...

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    I've tested samples using this kit.

    Because the background is so high, I end up getting a negative value, which is wrong, because previously I got positive values for the same samples.

    Thanks for your email.

    It does seem that this kit has an unusually high background. If you have purchased this kit within the last 6 months or so I would be happy to send you a replacement kit. If you are interested, please pass along the Abcam ...

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    I ordered two alpha ketoglutarate (akg) assay kits (ab83431).

    Both of them are from Lot GR77216-3.

    Both of these kits have a problem where the OD at 570 nm appears to be way too high for the alpha ketoglutarate standard curve. <...

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    This is a bit odd as we have done multiple QCs on this lot and have always got good results.


    The data does not look very bad, just that the background is quite high for some reason. If you look at the 10 nmol standard reading, you get a va...

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    Thank you very much for your reply,
    Im in a conference now and I dont have the ordering number with me. Can you please send me a replacement one?

    I found your order using the address provided in your signature and as requested, I have issued a free of charge replacement with the order number *****.

    To check the status of the order please contact our Customer Service team and reference this...

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    I had ordered alpha ketoglutarate assay kits (ab83431 – Lot: GR77216-2) last month and unfortunately the kits didn’t work. I started with the standard curve preparation and the results are as follow:

    As you see the zero value is very ...

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    Thank you forcontacting us. I am sorry to hear this kit is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

    Having...

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    I believe you have received the PO number. Looking forward to receive the standard very soon.
    Thanks,

    Thank you again for your emails, and I sincerely apologize for the delay in getting this replacement to you.

    The new standards arrived from the lab today, and I'm sending a free of charge kit ab83431 on the order ***. This will ship out tonight ...

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    I look forward to receiving the new standard component tomorrow.
    I believe you have received the PO number. Looking forward to receive the standard very soon.

    Thank you for your emails today and again for your patience with this situation. I have now found your original order, which can be used to send the replacement kit.

    I sincerely apologize, but there was a mistake in the shipment last week and we...

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    The protocol says, "Tissue (20 mg) or cells (2 x 10^6) are rapidly homogenized with 100 μl of ice cold PBS or other buffer (pH 6.5 - 8)." I know this means using a Dounce homogenizer, but I'm using T cells and I'm worried this won't lyse effe...

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    Thank you for your inquiry.

    Yes, buffers with pH range 6.5 – 8 can be used. Just make sure that the RIPA buffer is within the recommendedpH.
    I hope this information helps. Please contact us with any other questions.

    Standard came as a liquid and it should be lyophilized.

    Thank you for your call last week and for letting us know about the problem with this kit.

    We have received new standard components from the lab, and I will send you a free of charge replacement kit. Can you please confirm the PO#, order number,...

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