DIFFUSION BLOTTING FOR RAPID PRODUCTION OF MULTIPLE IDENTICAL IMPRINTS FROM SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS ON A SOLID SUPPORT

A simple and fast method for diffusion blotting of proteins from preca st SDS-PAGE gels (0.5 mm) was developed. The efficiency of protein tra nsfer was evaluated using C-14 labelled proteins. Diffusion blotting f or three minutes, gave a transfer of 10% compared to electroblotting. By doubling the transfer time for each subsequent imprint, four imprin ts were made from the same lane with similar amounts of protein transf erred onto each imprint. With a transfer time of three minutes each, i t was possible to obtain at least ten imprints with all the proteins v isible in all the imprints. There was no detectable loss in resolution as compared to electroblotting. The method also works well with an im mune-detecting system. The number of imprints which can be obtained, i s dependent on the sensitivity of the detection system and the amount of protein applied. The greatest advantage of diffusion blotting compa red to electroblotting is that several imprints can be made from each lane, and different antisera can be tested on identical imprints. The gel remains an its plastic support which prevents it from stretching a nd compression; this ensures identical imprints and facilitates more r eliable molecular mass determination. If only a few imprints are made, sufficient protein remains within the gel for general protein stainin g. These advantages make diffusion blotting the method of choice when quantitative protein transfer is not required. (C) 1998 Elsevier Scien ce B.V. All rights reserved.