A Short Review of Techniques for the Localisation of γ-Glutamyl Transferase Isoenzymes after Electrophoresis

Abstract

Summary The development of a sensitive method for the localisation of γ-glutamyl transferase isoenzymes after electrophoresis on various media is described. Recommendations on final conditions are given for optimal concentrations of substrate, acceptor and buffer, and for pH of the reaction mixture. The reaction conditions are compared with other isoenzyme localisation techniques previously reported.

gamma-Glutamyl transferase isoenzymes in human bile.

“…Using both electrophoresis procedures, the yGT isoenzymes were visualised by means of incubation with y-L-glutamylk-naphthylamide, coupling the liberated (x-naphthylamine with Fast Blue B according to the optimised procedure of Wenham et al (1978). More accurate quantitation of the isoenzyme fractions after polyacrylamide gel electrophoresis was accomplished by cutting the gel into 05-cm segments and incubating the homogenized gel segments in buffered substrate according to the method of Rosalki et al (1970).…”

Purification and Characterization of Gamma-Glutamyl Transpeptidase from Human Pancreas

“…The location of enzyme activity on the gel was visual ized by incubating with y-L-glutamyl-a-naphthylamide and Gly-Gly followed by coupling with fast blue B salt by the method of Whenham et al [9]. The protein was stained with Coomassie brillant blue R.…”

Serum gamma-glutamyltransferase isoenzymes in extrahepatic biliary obstruction.

“…The yGT isoenzymes fractionated by these two techniques were visualised by means of incubation with y-L-glutamyl ct-naphthylamide, coupling the liberated cx-naphthylamine with Fast Blue B, by the optimised method of Wenham et al (1978b). The isoenzymes were also quantitated after polyacrylamide gel electrophoresis by cutting the gel into 0 5 cm segments and incubating the homogenised gel segments in buffered substrate according to the method of Rosalki et al (1970).…”