[Cyanogen bromide peptide mapping of collagen polypeptides after separation using polyacrylamide gel electrophoresis...

Influence of various procedures for detection of collagen chains after electrophoresis in polyacrylamide gel on efficiency of their subsequent electroelution from the gel slices into gel of the second direction was studied. Staining of the gels with Coumassy R-250 under mild conditions or radioautography of the gels dried after electrophoresis decreased only slightly the electroelution efficiency as compared with the untreated gels. This observation suggests that direct procedures, which simplified distinctly the experiments, might be used for detection of the polypeptides in initial gel when peptide mapping was carried out using degradation of collagens by means of cyanogen bromide immediately in the gel slices. The procedures were developed, which enabled to perform mapping of collagen individual chains both labelled with 14C-amino acids and unlabelled fractions after their electrophoresis in polyacrylamide gel and the direct detection of polypeptides.