Novel and sensitive activity stains on polyacrylamide gel of serine and threonine dehydratase and ornithine aminotransferase.
A sensitive activity stain for serine and threonine dehydratase is described. After electrophoresis on thin-layer polyacrylamide gel, a reaction mixture containing L-serine or L-threonine as a substrate, NADH, lactate dehydrogenase and agar was solidified on the gel. The oxidation of NADH was recorded by contact printing on photographic paper with UV light. A novel activity stain for ornithine aminotransferase in the gel is also described. A reaction mixture containing substrates, o-aminobenzaldehyde and agar was solidified on the gel. A yellow color developed in a reaction between pyrroline-5-carboxylate and o-aminobenzaldehyde. The photograph was taken by contact printing. Both of the methods are highly sensitive and can detect less than 0.01 unit (mumol product/min at 37 degrees C) of each enzyme.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D012695:L-Serine Dehydratase / Q000032:analysis*; D008297:Male; D009953:Ornithine-Oxo-Acid Transaminase / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D013913:Threonine Dehydratase / Q000032:analysis*; D000637:Transaminases / Q000032:analysis*
D000637:Transaminases; D009953:Ornithine-Oxo-Acid Transaminase; D012695:L-Serine Dehydratase; D013913:Threonine Dehydratase