Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4302

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Immunoblotting: Submerged 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

Transfer of proteins from a Tris/glycine SDS-polyacrylamide gel to a membrane using the submerged method is achieved by placing the gel next to a piece of nitrocellulose filter, submerging this sandwich in a large volume of transfer buffer in a transfer tank, and running current from one side of the transfer tank to another. The proteins are then eluted by transferring them from the gel onto the filter. If carefully assembled, this is an effective method to transfer many proteins to a membrane. It demands a slightly longer time to set up than the semi-dry transfer (Immunoblotting: Semi-dry Electrophoretic . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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