Overview

  • Product name
    Branched Chain Amino Acid Assay Kit
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • Assay type
    Quantitative
  • Sensitivity
    > 0.2 nmol/well
  • Assay time
    0h 40m
  • Product overview

    Abcam's Branched Chain Amino Acid Assay Kit provides a simple convenient means of measuring the BCAA's in a variety of biological samples. The kit utilizes an enzyme assay in which BCAA is oxidatively deaminated, producing NADH which reduces the probe, generating a colored product (?max = 450 nm). Abcam's Branched Chain Amino Acid Assay Kit measures BCAA's in the range of 0 to 10 nmol per sample with a detection limit of ~0.2 nmol (~10 µM BCAA in sample). BCAA's are present in serum ~ 0.1-0.4 mM each (~0.125-1.5 mM combined).
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    The branched-chain amino acids or BCAA's, refer to the amino acids with non-linear aliphatic side-chains, namely leucine, isoleucine and valine. These three essential amino acids make up approximately 1/3 of skeletal muscle in the human body. BCAA's are currently used clinically to aid in the recovery of burn victims, as well as for strength supplementation for athletes. BCAA's, primarily Leu, can stimulate insulin secretion. The BCAA's have also been implicated in a wide range of other physiological effects.

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    BCAA Assay Buffer WM 1 x 25ml
    BCAA Enzyme Mix (lyophilized) Green 1 vial
    Leu Standard (1 µmol) Yellow 1 x 100µl
    WST Substrate Mix (lyophilized) Red 1 vial
  • Research areas
  • Relevance
    The branched-chain amino acids or BCAA’s, refer to the amino acids with non-linear aliphatic side-chains, namely leucine, isoleucine and valine. These three essential amino acids make up approximately 1/3 of skeletal muscle in the human body. BCAA’s are currently used clinically to aid in the recovery of burn victims, as well as for strength supplementation for athletes. BCAA’s, primarily Leu, can stimulate insulin secretion. The BCAA’s have also been implicated in a wide range of other physiological effects.
  • Alternative names
    • BCAA
    • Isoleucine
    • Leucine
    • Valine

Images

  • BRCAA levels measured in mouse tissue lysates (mg of extracted protein; background signal subtracted, mean of duplicates; +/- SD).

  • BRCAA levels measured in cell lysates (background signal subtracted, mean of duplicates; +/- SD).

  • BRCAA levels measured in human biological fluids (background signal subtracted, mean of duplicates; +/- SD).

  • Leucine Standard Curve performed according to the attached protocol.

Protocols

References

This product has been referenced in:
  • Serralde-Zúñiga AE  et al. Omental adipose tissue gene expression, gene variants, branched-chain amino acids, and their relationship with metabolic syndrome and insulin resistance in humans. Genes Nutr 9:431 (2014). Read more (PubMed: 25260659) »
See 1 Publication for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Question
Answer

The plasma samples can be diluted with water and then after adding to the well, the volume must be filled up to 50 ul with assay buffer.

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Answer


ab83374 (Branched Chain Amino Acid Assay Kit). We have not tested this kit with cell or tissue lysates yet but only with fluids like Cell culture supernatant, Urine, Serum, Plasma and can therefore not provide more than a general protocol below.

ab65347 (L-Amino Acid Assay Kit) has not been tested with cell lysates but the 96 well format is for liquid samples. Please use 50 ul samples per well. There is not need to grow the cells in the small wells.

The following general guidelines can be followed:

For cell samples: Start with ˜2X106 cells, suspend the cell pellet 500 μl (or ˜4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

For tissue samples: Start with 10-100 mg of the tissue, add 500-1000 μl (or ˜4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

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Answer

Thank you for contacting us.

This kit will detect all branched chain amino acids, regardless of whether they are single amino acids or peptides.

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

Merci de nous avoir contactés.

Le laboratoire m'a confirmé que nous n'avons malheureusement pas de données expérimentales pour ces kits sur des lysats musculaires.

Désolé de ne pas pouvoir vous aider davantage. N'hésitez pas néanmoins à nous contacter de nouveau si vous avez d'autres questions.

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Answer

Thank you for your inquiry.

I am happy to confirm that this kit will measure all the Branched chain amino acids (BCAAs).

All BCAAs will be deaminated to produce NADH, which is what this kit will actually measure.
The Leucine in the standard will also be converted to NADH. Therefore it does not matter which Branched chain amino acid is used as standard, it will still be measured in form of NADH.
Hope this information has been useful for you. Please let me know if you have any other questions.

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

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