Direct blotting, sequencing and immunodetection of proteins after five- minute staining of SDS and SDS-treated IEF gels with Nile red

The non-covalem dye Nile red allows the fast and simple fluorescent staining of protein bands in sodium dodecyl sulfate (SDS)-polyacrylamide gels. This procedure has been extended to polyacrylamide isoelectric focusing gels than do not contain SDS. Unlike the current methods using Cuomassie blue or silver for gel staining. Nile red staining does not preclude the direct electroblotting of protein bands onto polyvinylidene difluoride membranes, and the transferred proteins can be used directly for immunoblotting analysis and for N-terminal microsequencing.