Protocol

Preparation of Vertical SDS Slab Gels: Simultaneous Casting of Multiple-Gradient Gels

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

Gradient SDS-PAGE gels provide the best resolution over a wide range of molecular weights, resulting in sharper protein spots, because diffusion is minimized by the decreasing pore size in the gel. However, gradient gels are more difficult to produce reproducibly; thus, they are commonly cast with multiple-gel casters, which allows for an identical set of gels to be produced for an experiment. Presented here is a method for casting gradient gels using a multiple-gel casting system.