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

ABSTRACT: 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.

SUBMITTER: Domanski M

PROVIDER: S-EPMC3605737 | biostudies-other | 2012 May

REPOSITORIES: biostudies-other

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

Domanski Michal M Molloy Kelly K Jiang Hua H Chait Brian T BT Rout Michael P MP Jensen Torben Heick TH LaCava John J

BioTechniques 20120501 0

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. ...[more]

PMID: 22668517

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