Tandem affinity purification of functional TAP-tagged proteins from human cells
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
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.
Details
Originalsprache | Englisch |
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Seiten (von - bis) | 1145-1151 |
Seitenumfang | 7 |
Fachzeitschrift | Nature protocols |
Jahrgang | 2 |
Ausgabenummer | 5 |
Publikationsstatus | Veröffentlicht - 2007 |
Peer-Review-Status | Ja |
Externe IDs
PubMedCentral | PMC2957861 |
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Scopus | 34250155598 |
Schlagworte
Schlagwörter
- Animals, Cell Cycle Proteins/genetics, Chromatography, Affinity/methods, Chromosomes, Artificial, Bacterial/genetics, Electrophoresis, Polyacrylamide Gel, HeLa Cells, Humans, Mass Spectrometry, Mice, Multiprotein Complexes/isolation & purification, RNA Interference