Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4570

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In Situ SDS-PAGE Peptide-Mapping Procedure (Cleveland Method)

Richard J. Simpson

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

In this protocol, sample proteins are first separated by either one- or two-dimensional gel electrophoresis. The gel is briefly stained and destained, and the proteins are excised for peptide mapping. Peptide fragmentation (either enzymatically or chemically) is performed in situ in the gel pieces, and the partial digest is separated in a second gel.


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