Protocol

Immunohistochemistry on Sections of Plant Tissues Using Alkaline-Phosphatase-Coupled Secondary Antibody

This protocol was adapted from “How to Study Gene Expression,” Chapter 7, in Arabidopsis (eds. Weigel and Glazebrook). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2002.

INTRODUCTION

Like RNA, proteins can be localized in situ on tissue sections. In situ localization or immunohistochemistry provides information on the cellular and subcellular distribution of proteins in different tissues. This protocol describes an alternative to the use of ABC (a preformed Avidin:Biotinylated enzyme Complex used in conjunction with a biotinylated secondary antibody) for non-fluorescent detection in immunohistochemistry. Here, an alkaline-phosphatase-coupled secondary antibody is used with colorimetric substrates for antigen detection.

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