UNIT 14.6 Immunohistochemistry

  1. Simon Watkins

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1406s31

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Watkins, S. 2001. Immunohistochemistry. Current Protocols in Molecular Biology. 14:14.6.

Author Information

  1. Harvard Medical School and Dana-Farber Cancer Institute, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 1995

This is not the most recent version of the article. View current version (1 JAN 2008)


This unit surveys immunohistochemical techniques for protein localization. The first four protocols focus on indirect immunofluorescence which is the easiest and most widely used method of optical detection. Additional protocols present methods suitable for bright-field microscopy, such as immunoperoxidase and immunogold labeling. The commentary contains an extensive troubleshooting guide with a list of suitable routine controls. The chosen method of tissue or cellular preparation must be suitable to answer the fundamental objective of the protocol (i.e., where the protein is localized within the system). Furthermore, the choice of fluorochrome and the method of labeling are also important to optimize the sensitivity of antigen localization. The basic protocol is therefore divided into two sections: (1) cellular preparations (monolayers of adherent cells and cells grown in suspension) and (2) tissue section labeling.