Assay of long-chain acyl-CoAs in a complex reaction mixture.
A method has been developed which allows the quantitative analysis of labeled or unlabeled acyl-CoAs in complex reaction mixtures. The method is based on (a) a quantitative solubilization of acyl-CoAs and lipids, directly in the reaction vessel, by 0.05 M Tris-HCl, pH 7.5/CHCl3/CH3OH (1/3/3, v/v/v); (b) monodimensional TLC of aliquots of the whole reaction mixture, resolving malonyl-CoA, acetyl-CoA, long-chain acyl-CoAs, polar lipids and neutral lipids plus free fatty acids; and (c) quantitation by TLC densitometry and/or TLC radiochromatography. All fractions--and particularly long chain-acyl-CoAs--can then be analyzed for distribution and label of fatty acyl moieties.
D000214:Acyl Coenzyme A / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D002855:Chromatography, Thin Layer; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000032:analysis*; D008861:Microsomes / Q000378:metabolism; D010944:Plants / Q000378:metabolism; D012995:Solubility
D000214:Acyl Coenzyme A; D008055:Lipids