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Affinity filtration coupled with capillary-based affinity purification for the isolation of protein complexes
The isolation of complex macromolecular assemblies at the concentrations required for structural analysis represents a major experimental challenge. Here we present a method that combines the genetic power of site-specific recombination in order to selectively “tag” one or more components of a prote...
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| Published in: | Analytical biochemistry 2013-08, Vol.439 (1), p.47-49 |
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| Main Authors: | , , , , , , , , |
| Format: | Article |
| Language: | English |
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| cites | cdi_FETCH-LOGICAL-c374t-a5b2cbbf84ce32758a86934d2cff5b969d1cbff54338a2e9166878bbe3f1e3c03 |
| container_end_page | 49 |
| container_issue | 1 |
| container_start_page | 47 |
| container_title | Analytical biochemistry |
| container_volume | 439 |
| creator | Qureshi, M.S. Sheikh, Q.I. Hill, R. Brown, P.E. Dickman, M.J. Tzokov, S.B. Rice, D.W. Gjerde, D.T. Hornby, D.P. |
| description | The isolation of complex macromolecular assemblies at the concentrations required for structural analysis represents a major experimental challenge. Here we present a method that combines the genetic power of site-specific recombination in order to selectively “tag” one or more components of a protein complex with affinity-based rapid filtration and a final step of capillary-based enrichment. This modified form of tandem affinity purification produces highly purified protein complexes at high concentrations in a highly efficient manner. The application of the method is demonstrated for the yeast Arp2/3 heptameric protein complex involved in mediating reorganization of the actin cytoskeleton. |
| doi_str_mv | 10.1016/j.ab.2013.03.035 |
| format | article |
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| identifier | ISSN: 0003-2697 |
| ispartof | Analytical biochemistry, 2013-08, Vol.439 (1), p.47-49 |
| issn | 0003-2697 1096-0309 |
| language | eng |
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| source | ScienceDirect Journals |
| subjects | Actin-Related Protein 2-3 Complex - isolation & purification Affinity Arp2/3 Capillary Chromatography, Affinity - methods complexing filtration Filtration - methods microfilaments Protein complex Protein purification Proteins - isolation & purification Saccharomyces cerevisiae Proteins - isolation & purification site-specific recombination yeasts |
| title | Affinity filtration coupled with capillary-based affinity purification for the isolation of protein complexes |
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