Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels

摘要: Coomassie blue-stained, heat-dried, and computer-imaged two-dimensional gels used to develop comprehensive human protein data bases served as the source generate partial amino acid sequences. The spots were collected from multiple gels, rehydrated, concentrated by stacking into a new gel, electroblotted onto inert membranes, in situ-digested with trypsin. Peptides eluting membranes separated HPLC sequenced. Using this procedure, it was possible sequences 13 proteins recorded amnion cell base. Eight of these matched those stored bases, demonstrating that systematic analysis computerized gel electrophoresis can be directly linked microsequencing methods. latter technique offers unique opportunity link information contained derived forthcoming DNA sequence on genome.