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

Biotechniques. 2012 May;0(0):1-6. doi: 10.2144/000113864.


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.