Branched Chain Amino Acid Assay Kit / BCAA Assay Kit (ab83374)

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.

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.

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.

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.

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.