Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4615

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Detection of Ubiquitylated Proteins in Yeast

William P. Tansey

Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA

Corresponding author (tansey{at}cshl.edu)


INTRODUCTION

This assay is performed to detect ubiquitylated proteins in yeast. Yeast that have been transformed with a vector expressing polyhistidine-tagged ubiquitin (Ub) under the control of a copper-inducible promoter are grown, induced with copper, and harvested. Total ubiquitylated proteins are then recovered by nickel-affinity chromatography, and specific proteins are detected by Western blotting.


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