Protocol

SDS-PAGE Peptide-Mapping Procedure (Cleveland Method)

This protocol was adapted from "Peptide Mapping and Sequence Analysis of Gel-Resolved Proteins," Chapter 7, in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

INTRODUCTION

The Cleveland method for peptide mapping of proteins relies on partial hydrolysis of the protein yielding a considerable number of large peptides that can be readily identified by SDS-PAGE or tricine-SDS-PAGE. Small peptides obtained with a total digest are often poorly fixed within the gel and usually contribute nothing to the peptide map. In the method described here, the protein is partially digested in solution, and the resulting peptide fragments are separated by SDS-PAGE.