Purification and radioimmunoassay of rat lactate dehydrogenase A and B subunits.
We have developed procedures for purifying lactate dehydrogenase isoenzymes from rat tissues that involve two affinity-chromatography steps and that facilitate the isolation of milligram quantities of highly purified proteins within 2--3 days. Antibodies raised against pure A and B subunits in rabbits and hens were used in radioimmunoassays and showed negligible cross-reactivity with heterologous subunits. The radioimmunoassays provide a sensitive method for measuring nanogram amounts of A-subunit and B-subunit polypeptides in tissue homogenates and were employed to characterize the enzyme purification procedures.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D011919:Rats, Inbred Strains
D007106:Immune Sera; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase