Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4523

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Antibody Addition and Detection for Staining Caenorhabditis elegans

Ed Harlow and David Lane

This protocol was adapted from "Staining Tissues," Chapter 6, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.


INTRODUCTION

The most common method of detection for staining worms is to use fluorochrome-labeled reagents, because of the desire for high resolution of the antigen location. Extreme care should be used to identify and verify positive reactions, however, because cross-reactions are common. Counterstaining is essential for examining worms by immunofluorescence and is used to identify the exact cell in which an antigen appears. Methods for counterstaining include labeling all cells with a fluorescent dye that is specific for nucleic acids (e.g., DAPI or propidium iodide) and using GFP driven by tissue-specific promoters.


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