Laney JD and Hochstrasser M (2011) Analysis of protein ubiquitination. Curr Protoc Protein Sci Chapter 14():Unit14.5
Abstract: Attachment of ubiquitin (Ub) to a protein requires a series of enzymes that recognize the substrate and promote Ub transfer. Several methods are described in this unit for determining if a protein has Ub-transferring activity. They include immunoblotting of immunoprecipitated proteins, affinity purification using His-tagged Ub, assaying for auto-ubiquitination of E3, and assaying ubiquitination of a model substrate protein in vitro and in E. coli cells that express Ub-ligation enzymes. These methods are suitable for a variety of eukaryotic cells, but techniques are specifically described for use with yeast and mammalian cells. Curr. Protoc. Protein Sci. 66:14.5.1-14.5.13. (c) 2011 by John Wiley & Sons, Inc.
|Status: Published||Type: Journal Article||PubMed ID: 22045559|
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