A method for branched-chain amino acid aminotransferase activity in microgram and nanogram tissue samples
✍ Scribed by Carol S. Hintz; William R. Turk; Natalie Cambon; Helen B. Burch; Patti M. Nemeth; Oliver H. Lowry
- Publisher
- Elsevier Science
- Year
- 1985
- Tongue
- English
- Weight
- 399 KB
- Volume
- 146
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A method for branched-chain amino acid aminotransferase is described which is based on running the reaction in the reverse of the usual direction with glutamate and r*-ketoisocaproate as substrates. The cY-ketoglutarate generated is reduced with glutamate dehydrogenase and NADH. For sensitivity in the nanomole range, the NAD+ generated is measured directly by converting to the highly fluorescent strong alkali product. For smaller samples, down to the 0.2-to 2-pmol range, the NAD+ is amplified by enzymatic cycling. 0 1985 Academic PW.S. IX.