Improved methodology for the affinity isolation of human protein complexes expressed at near endogenous levels.
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.
Centre for mRNP Biogenesis and Metabolism, Department of Molecular Biology and Genetics, Aarhus University, Aarhus C, Denmark.
Michal
M
Domanski
Kelly
K
Molloy
Hua
H
Jiang
Brian T
BT
Chait
Michael P
MP
Rout
Torben Heick
TH
Jensen
John
J
LaCava