Part of the Methods in Molecular Biology™ book series (MIMB, volume 105)
Immunohistochemistry and Immunocytochemistry
Immunohistochemistry and immunocytochemistry are powerful techniques for localizing the molecular expression of proteins in tissues and cells, especially when combined with the in situ hybridization technique. Immunohistochemistry and immunocytochemistry techniques are primarily considered to be qualitative measurements, but when used together with a computerized imaging program, staining distribution and intensity can be semiquantified. This chapter describes immunochemical techniques routinely used in our laboratories for protein localization in tissue sections and cultured cells with the representative staining shown in Figs. 1 and 2. The protocols are based on an indirect immunoperoxidase detection via the avidin:biotinylated-peroxidase complex method with 3,3′-diaminobenzidine as the chromogen, a popular method suitable for bright-field microscopy. The first immunohistochemical protocol described is intended for paraffin-embedded tissue sections and the second immunocytochemical protocol is intended for cultured cells. It is highly recommended that, before beginning the protocol, the operator read Subheading 4.
KeywordsTissue Block Sodium Cacodylate Sodium Cacodylate Buffer Clearing Agent Chlorine Bleach
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© Humana Press Inc., Totowa, NJ 1998