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Improved methodology for the affinity isolation of human protein complexes expressed at near endogenous levels

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  • Michal Domanski, Danmark
  • Kelly Molloy, Laboratory Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, USA
  • Hua Jiang, Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, USA
  • Brian T Chait, Laboratory Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, USA
  • Michael P Rout, Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, USA
  • Torben Heick Jensen
  • John LaCava, Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, USA

An efficient and reliable procedure for the capture of affinity-tagged proteins and associated complexes from human cell lines is reported. Through multiple optimizations, high yield and low background affinity-purifications are achieved from modest quantities of human cells expressing endogenous-level tagged proteins. Isolations of triple-FLAG and GFP-tagged fusion proteins involved in RNA metabolism are presented.

OriginalsprogEngelsk
TidsskriftBioTechniques
Vol/bind0
Nummer0
Sider (fra-til)1-6
Antal sider6
ISSN0736-6205
DOI
StatusUdgivet - maj 2012

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