Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4559

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Preparative 2D Gel Electrophoresis with Immobilized pH Gradients: SDS-PAGE of Proteins

Wayne R. Stochaj, Tom Berkelman, and Nancy Laird

This protocol was adapted from "Preparative 2D Gel Electrophoresis with Immobilized pH Gradients," Chapter 4 in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.


INTRODUCTION

Following first-dimension IEF and equilibration of the IPG gel strips, the proteins are separated on the basis of their molecular weight in the second dimension on an SDS-PAGE gel. Systems for this separation are available from a variety of suppliers and are commonly found in many protein chemistry laboratories. This protocol describes a method for placement of the IPG strip and gives some recommended electrophoresis conditions for these second-dimension gels.


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