Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4278

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Epitope Mapping Using Synthetic Biotin-Labeled Peptides

Ed Harlow and David Lane

This protocol was adapted from "Epitope Mapping," Chapter 11, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

The first 100 words of the full text of this article appear below.


INTRODUCTION

A set of overlapping synthetic peptides is synthesized, each corresponding to a small segment of the linear sequence of a protein antigen and arrayed on a solid phase. The panel of solid-phase peptides is then probed with a test antibody, and bound antibody is detected using an enzyme-labeled secondary antibody. This method is very rapid and can be extraordinarily successful, usually yielding unequivocal results with antibodies that work well in immunoblotting and that recognize denaturation-resistant epitopes. It does not work with antibodies that only react with nondenatured native protein.


MATERIALS

Reagents

caution Biotin-labeled peptides

If time and money were not constraints, the optimal . . . [Full Text of this Article]

Equipment


METHOD


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