Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4344

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Immunofluorescence Detection of RNAi-Induced Protein Knockdown in Mammalian Cells

Thomas Tuschl

This protocol was adapted from "Mammalian RNA Interference," contributed by Thomas Tuschl, Chapter 13, in RNAi: A Guide to Gene Silencing (ed. Hannon). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes the use of a specific antibody that recognizes the targeted gene product to detect RNAi-induced gene knockdown in mammalian cells. Western blot technology can be used as an alternative (see Detection of RNAi-Induced Protein Knockdown in Mammalian Cells by Western Blotting).


MATERIALS

Reagents

Cells from Tranfection of Mammalian Cells with siRNA Duplexes grown on coverslips

caution Hoechst 33342 (bisbenzimide; 15091)

caution Methanol, chilled to –10ºC

Moviol mounting medium (Hoechst, www.hoechst.com)

recipe Phosphate-buffered saline (PBS) (pH 7.1)

. . . [Full Text of this Article]

Equipment


METHOD


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