Staining of argininosuccinate lyase activity in polyacrylamide gel.
A procedure for the direct staining of argininosuccinate lyase activity in polyacrylamide gel is described. The method was based on coupling one of the enzymatic products fumarate with fumarase and malic enzyme catalyzed reactions. Fumarate was first converted to L-malate by fumarase. Malic enzyme then catalyzed the oxidative decarboxylation of L-malate to give CO2 and pyruvate with concomitant reduction of NADP+ to NADPH. Finally the reducing power of NADPH was coupled to phenazine methosulfate and in turn to nitroblue tetrazolium yielding a deeply colored insoluble formazan which may be quantitized or semiquantitized by densitometer.
Lee
H J
HJ
Department of Biochemistry, National Defence Medical Center, Taipei, Taiwan, R.O.C.
Chiou
S H
SH
Chang
G G
GG
D000818:Animals; D001123:Argininosuccinate Lyase / Q000032:analysis*; D003459:Crystallins / Q000737:chemistry*; D004372:Ducks; D004591:Electrophoresis, Polyacrylamide Gel; D013194:Staining and Labeling
D003459:Crystallins; D001123:Argininosuccinate Lyase