File:Schematic depiction of two affinity purification approaches..jpg
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English: A. Tandem affinity chromatography: a protein of interest is first engineered to contain a Protein A (filled in circle) tag, a Tobacco Echo Virus recognition site (TEV, filled in triangle), and a calmodulin binding peptide (CBP) tag (filled in ellipse). Extracts are made from cells expressing the tagged protein, which should contain associated proteins and contaminants. These complexes are bound to an IgG column, and washed to remove majority of contaminants. TEV protease is then used to elute the semi-purified protein complexes which are subsequently absorbed onto a calmodulin column. After further washing, purified protein complexes containing the protein of interest and its associated proteins are eluted by calcium chelation (EGTA) and identified using LC-MS/MS. |
| Date | Published July 14, 2008. |
| Source |
[1] Direct
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| Author | Wang, J., Trowbridge, J.J., Rao, S. and Orkin, S.H., Proteomic studies of stem cells (July 14, 2008), StemBook, ed. The Stem Cell Research Community, StemBook, doi/10.3824/stembook.1.4.1, http://www.stembook.org. |
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  This file is licensed under the Creative Commons Attribution 3.0 Unported license.
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| Date/Time | Thumbnail | Dimensions | User | Comment | |
|---|---|---|---|---|---|
| current | 19:04, 5 April 2013 | | 563 × 600 (93 KB) | Smallbot (talk | contribs) | Uploading CC-BY images from the the StemBook http://www.stembook.org/ 151/173 |
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