Tandem affinity purification of functional TAP-tagged proteins from human cells
Open Access
- 3 May 2007
- journal article
- research article
- Published by Springer Nature in Nature Protocols
- Vol. 2 (5) , 1145-1151
- https://doi.org/10.1038/nprot.2007.172
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.Keywords
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