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Journal Article

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

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Poser,  Ina
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Buchholz,  Frank
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Gregan, J., Riedel, C. G., Petronczki, M., Cipak, L., Rumpf, C., Poser, I., et al. (2007). Tandem affinity purification of functional TAP-tagged proteins from human cells. Nature Protocols, 2(5), 1145-1151.


Cite as: https://hdl.handle.net/21.11116/0000-0001-0FA8-2
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.