The Art of IHC: Immunohistochemistry with Anti-ACTIVE^® Antibodies

Introduction

By Randy Hoffman
Promega Corporation

Introduction

This image represents staining of CaM KII activated neurons by Anti-ACTIVE^® CaM KII pAb in an adult rat brain. The image rendered here was obtained by subjecting the original image to a texture filter in Photoshop (Adobe).

Protein expression, as monitored by in situ cellular and subcellular localization, has become exceedingly important in the post genome sequencing era. Immunohistochemistry, coupled with traditional molecular and biochemical techniques, continues to be a powerful tool for studying gene expression. By utilizing robust antibodies capable of detecting antigens in fixed tissue sections, scientists can delineate gene expression patterns under various experimental conditions. Promega has developed many polyclonal antibodies useful for the study of eukaryotic signal transduction. Anti-ACTIVE^® MAPK, JNK/p38 and CaM KII antibodies are affinity-purified rabbit polyclonal antibodies that are specific for the active, phosphorylated forms of these kinases. In addition, Promega has developed polyclonal antibodies useful for the study of apoptosis. Anti-PARP p85 Fragment pAb, Anti-ACTIVE^® Caspase-3 pAb, and Anti-pS⁴⁷³ Akt pAb are affinity purified rabbit polyclonal antibodies specific for caspase-cleaved PARP, the active form of caspase-3, and phosphorylated Akt, respectively.

The primary purpose of this article is to demonstrate the robust staining characteristics of these antibodies in fixed tissue sections from mouse and rat, but a secondary purpose is the demonstration of IHC as an artform.