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

by: J Gregan, CG Riedel, M Petronczki, L Cipak, C Rumpf, I Poser, F Buchholz, K Mechtler, K Nasmyth

Nat Protoc, Vol. 2, No. 5. (2007), pp. 1145-1151.

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Abstract

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.

@article{citeulike:1638355, abstract = {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.}, address = {Department of Chromosome Biology, Max F. Perutz Laboratories, University of Vienna, Dr Bohr-Gasse 1, 1030 Vienna, Austria. juraj.gregan@univie.ac.at}, author = {Gregan, J. and Riedel, C. G. and Petronczki, M. and Cipak, L. and Rumpf, C. and Poser, I. and Buchholz, F. and Mechtler, K. and Nasmyth, K. }, citeulike-article-id = {1638355}, doi = {http://dx.doi.org/10.1038/nprot.2007.172}, issn = {1750-2799}, journal = {Nat Protoc}, number = {5}, pages = {1145--1151}, posted-at = {2007-09-09 09:21:04}, priority = {2}, title = {Tandem affinity purification of functional TAP-tagged proteins from human cells.}, url = {http://dx.doi.org/10.1038/nprot.2007.172}, volume = {2}, year = {2007} }

RIS record

TY - JOUR ID - citeulike:1638355 L3 - citeulike-article-id:1638355 TI - Tandem affinity purification of functional TAP-tagged proteins from human cells. JF - Nat Protoc VL - 2 IS - 5 SP - 1145 EP - 1151 AD - Department of Chromosome Biology, Max F. Perutz Laboratories, University of Vienna, Dr Bohr-Gasse 1, 1030 Vienna, Austria. juraj.gregan@univie.ac.at SN - 1750-2799 N2 - 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. AU - Gregan, J AU - Riedel, CG AU - Petronczki, M AU - Cipak, L AU - Rumpf, C AU - Poser, I AU - Buchholz, F AU - Mechtler, K AU - Nasmyth, K PY - 2007/// UR - http://dx.doi.org/10.1038/nprot.2007.172 ER -

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