A method for branched-chain amino acid aminotransferase activity in microgram and nanogram tissue samples.
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 alpha-ketoisocaproate as substrates. The alpha-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.
D000818:Animals; D005612:Freeze Drying; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008832:Microchemistry; D009132:Muscles / Q000201:enzymology; D011817:Rabbits; D051381:Rats; D000637:Transaminases / Q000032:analysis*
D000637:Transaminases; C019798:branched-chain-amino-acid transaminase