Sequence Analysis of the NH2-Terminally Blocked Proteins Immobilized on PVDF Membranes from Polyacrylamide Gels

Abstract

In 1985, a new protein preparation method for microsequencing was established (Vandekerckhove et al. 1985), in which sub-nanomole amounts of proteins were separated by one-dimensional or two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) and then transferred from the gel onto a polybrene-treated glass fiber filter by electroblotting and sequenced directly by a gas-phase sequencer. Two years later, it was found that a polyvinylidene difluoride (PVDF) membrane could be used in place of the glass fiber filter for efficient electroblotting and gas-phase sequencing (Matsudaira 1987). The PVDF membrane filter has several advantages over the glass fiber filter in, for example, protein binding capacity, protein detection on the filter, and handling. Since 1987, electroblotting/sequenc-ing using the PVDF membrane has come to be widely used in protein sequence analysis.