Tandem affinity purification of functional TAP-tagged proteins from human cells

被引：44

作者：

Gregan, Juraj ^{[1
,2
]}

Riedel, Christian G. ^{[2
]}

Petronczki, Mark ^{[2
]}

Cipak, Lubos ^{[1
]}

Rumpf, Cornelia ^{[1
]}

Poser, Ina ^{[3
]}

Buchholz, Frank ^{[3
]}

Mechtler, Karl ^{[2
]}

Nasmyth, Kim ^{[2
,4
]}

机构：

[1] Univ Vienna, Dept Chrosome Biol, Max F Perutz Labs, A-1030 Vienna, Austria

[2] Res Inst Mol Pathol, A-1030 Vienna, Austria

[3] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany

[4] Univ Oxford, Dept Biochem, Oxford OX1 3QU, England

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 05期

基金：

奥地利科学基金会;

关键词：

D O I：

10.1038/nprot.2007.172

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Tandem affinity purification ( TAP) is a generic two-step affinity purification protocol for isolation of TAP-tagged proteins together with associated proteins. We used bacterial artificial chromosome to heterologously express TAP-tagged murine Sgo1 protein in human HeLa cells. This allowed us to test the functionality of the Sgo1-TAP protein by RNA interference-mediated depletion of the endogenous human Sgo1. Here, we present an optimized protocol for purification of TAP-tagged Sgo1 protein as well as KIAA1387 from HeLa cells with detailed instructions. The purification protocol can be completed in 1 day and it should be applicable to other proteins.

引用

页码：1145 / 1151

页数：7

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