Journal
NATURE PROTOCOLS
Volume 2, Issue 5, Pages 1145-1151Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.172
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Funding
- Austrian Science Fund FWF [P 18955] Funding Source: Medline
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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.
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