Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4167

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Yeast Immunofluorescence

David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern

This protocol was adapted from "Yeast Immunofluorescence," Techniques and Protocols 12, in Methods in Yeast Genetics, 2005 edition, by David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

The first 100 words of the full text of this article appear below.


INTRODUCTION

This protocol describes a method based on that of Pringle for immunofluorescent staining of yeast, as would be done in gene replacement experiments.


MATERIALS

Reagents

caution Acetone, cold in a coplin jar surrounded by dry ice

Begin cooling the acetone 1-2 hours before beginning the experiment

Appropriate yeast culture

recipe Block solution

caution Formaldehyde (EM grade, Polysciences), 10%

recipe KM solution

recipe KM solution containing 1.2 M sorbitol

caution Methanol, cold, in a coplin jar surrounded by dry ice

Begin cooling the methanol 1-2 hours before beginning the experiment

recipe caution Mounting solution

recipe Polylysine (>400,000 MW), 0.1%

Primary and secondary antibody conjugate diluted appropriately in block solution

caution Triton X-100 (optional)

. . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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