Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4529

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Unmasking Hidden Epitopes Using the Microwave Oven

Ed Harlow and David Lane

This protocol was adapted from "Staining Tissues," Chapter 6, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.


INTRODUCTION

Several methods have been developed to "retrieve" antigens that have been masked by fixation. The principle behind using the microwave oven method described here is to use extended periods of heat to break some of the subcellular structures that block antibody access. Be aware that any of the antigen retrieval methods should be avoided wherever possible, because they may introduce artifactual false-positive staining. Methods for antigen retrieval are specific for each antibody/antigen combination. No attempts to establish quantitative data following these methods should be considered.


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