| Literature DB >> 19107412 |
Helle D Ulrich1, Adelina A Davies.
Abstract
Ni-NTA affinity chromatography under denaturing conditions has proven to be a powerful method for the isolation of SUMO conjugates from total cell extracts, as it minimizes deconjugation and excludes noncovalent interactions. This chapter describes the use of both His-tagged SUMO and a His-tagged target protein for the characterization of the sumoylation process in the budding yeast Saccharomyces cerevisiae. Two well-studied model substrates, the septin Cdc3 and the replication clamp protein PCNA, are used as examples, but the protocol can easily be adapted to other targets and organisms.Entities:
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Year: 2009 PMID： 19107412 DOI： 10.1007/978-1-59745-566-4_6
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745