Protocol

Preparation of Immobilized Enzyme Microcolumns

This protocol was adapted from "The Use of Mass Spectrometry in Proteomics," Chapter 8, in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

INTRODUCTION

Peptide mapping by MALDI-TOF-MS has emerged as a powerful proteomics tool for identifying and characterizing proteins. One of the key steps in this method involves the proteolytic cleavage of proteins, followed by MS analysis of the generated peptides. A limitation of the method, especially for high-throughput proteomics, is the speed and efficiency of proteolytic cleavage. In this protocol, a method for rapid tryptic digestion using immobilized enzyme microcolumns is described.