Direct sequencing of peptides transferred onto polyvinylidene difluoride membrane from sodium dodecyl sulfate-polyacrylamide gel electrophoresis by electroblotting

Protein stained with coomassie brilliant blue (CBB) on polyacrylamide gels were digested with lysylendopeptidase in the presence of sodium dodecyl sul fate. The resulting peptides were then separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto a pol yvinylidene difluoride (PVDF) membrane. After CBB R-250 of the staining pep tide bands on the PVDF-blotted they were loaded directly into the sequencer . Using PVDF membrane for peptide separation and sequencing is quite simple and in contrast to separation of peptides by reversed-phase liquid chromat ography, it is not affected by sodium dodecyl sulfate. (C) 2001 Elsevier Sc ience B.V. All rights reserved.