Tandem affinity purification of functional TAP-tagged proteins from human cells
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
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
| Original language | English |
|---|---|
| Pages (from-to) | 1145-1151 |
| Number of pages | 7 |
| Journal | Nature protocols |
| Volume | 2 |
| Issue number | 5 |
| Publication status | Published - 2007 |
| Peer-reviewed | Yes |
External IDs
| PubMedCentral | PMC2957861 |
|---|---|
| Scopus | 34250155598 |
Keywords
Keywords
- 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