Immunoprecipitation and electrophoretic analysis of four human acyl-CoA dehydrogenases and electron transfer flavoprotein using antibodies raised against the corresponding rat enzymes.
We prepared monospecific antisera in rabbits against purified rat short-, medium-, and long-chain acyl-CoA dehydrogenases, isovaleryl-CoA dehydrogenase, and ETF and tested the immunocross-reactivity to the corresponding human enzymes. Each antiserum specifically reacted with the corresponding human enzyme. When immunoprecipitates were analyzed by SDS-PAGE, the mobilities of all the human acyl-CoA dehydrogenases and ETF subunits were identical to those of the rat counterparts with a single exception. Human medium-chain acyl-CoA dehydrogenase had a mobility on SDS-PAGE slightly slower than that of rat medium-chain acyl-CoA dehydrogenase, suggesting that human medium-chain acyl-CoA dehydrogenase was 1 kDa larger than the rat counterpart. The immunocross-reactivity of the antisera, raised against the rat acyl-CoA dehydrogenases and ETF to the human counterpart, provide useful tools for the study of mutant enzymes in cells from patients with a genetic defect of acyl-CoA dehydrogenases of ETF.
D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000302:isolation & purification*; D000906:Antibodies; D000936:Antigen-Antibody Complex; D044927:Electron-Transferring Flavoproteins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005420:Flavoproteins / Q000302:isolation & purification*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight
D000906:Antibodies; D000936:Antigen-Antibody Complex; D044927:Electron-Transferring Flavoproteins; D005420:Flavoproteins; D044942:Acyl-CoA Dehydrogenase, Long-Chain