Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4332

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Detection of Horseradish Peroxidase-Labeled Reagents with Diaminobenzidine/Metal

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.

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


INTRODUCTION

Horseradish peroxidase-linked secondary reagents are best used for low-resolution studies that are aimed at the rapid detection of the presence or localization of the antigen. The major advantages of using this detection method are the sensitivity and the ability to observe the results with just a light microscope. Extra sensitivity can be found by observing the precipitated enzyme products by interference reflection microscopy. Diaminobenzidine (DAB) is one of the most sensitive substrates for horseradish peroxidase. It yields an intense brown product that is insoluble in both water and alcohol. It can be made more sensitive by adding metal salts such . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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