Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4301

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Immunoblotting: Semi-Dry Electrophoretic Transfer of Proteins from Gels to Membranes

Ed Harlow and David Lane

This protocol was adapted from "Immunoblotting," Chapter 8, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

The first 100 words of the full text of this article appear below.


INTRODUCTION

The transfer of proteins from a Tris/glycine SDS-polyacrylamide gel to a membrane using a semi-dry method is achieved by placing the gel next to a piece of nitrocellulose filter. This sandwich is placed directly between two plate electrodes, and the proteins are then transferred from the gel onto the filter. Good contact between the electrodes, gel, and filter is maintained by using several sheets of pre-wetted glass-fiber filter paper on both the cathode and anode sides. A simple buffer system using diluted Tris/glycine SDS-polyacrylamide gel running buffer is used. This method gives even and rapid transfer and does not require . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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