Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4296

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Indirect Detection Using Horseradish Peroxidase-Labeled Reagents

Ed Harlow and David Lane

This protocol was adapted from "Staining Cells," Chapter 5, 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 the 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. Diaminobenzidine (DAB) is one of the most sensitive substrates for horseradish peroxidase, and it can be made more sensitive by adding metal salts such as cobalt or nickel to the substrate solution. The products are stable in both water and alcohol. For indirect detection using fluorochrome-labeled reagents, see Indirect Detection Using . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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