Determination of specific radioactivity of amino acids in proteins directly on polyacrylamide gels: An application to l-leucine

✍ Scribed by A.F. Martin; G. Prior; R. Zak

• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 528 KB
• Volume: 72
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90569-8

✦ Synopsis

A method has been developed for determination of the specific radioactivity of t-leucine in different proteins or protein subunits which have been separated by polyacrylamide gel electrophoresis. After electrophoresis, the proteins are visualized by staining with Coomassie Brilliant Blue R, and the desired bands are excised and hydrolyzed with hydrochloric acid. In one aliquot of the hydrolysate, the radioactivity is determined, and in another, the amino acid concentration. For this purpose, we have employed an isotope dilution procedure, using the aminoacyl synthetase reaction. With this technique, IOO-I500 picomol of a single amino acid can be detected.