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Question (75669) | Branched Chain Amino Acid Assay Kit (ab83374)

Inquiry: Can you please assit with more detailed sample prep instructions for assay kits: - L-Amino Acid Assay Kit ab65347 - Branch Chain Amino Acid Kit ab83374 The protocol booklet does not specify (with regards to cell lization/seeding in microtitre plate, etc?)


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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