Reference: Qureshi MS, et al. (2013) Affinity filtration coupled with capillary-based affinity purification for the isolation of protein complexes. Anal Biochem 439(1):47-9

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Abstract


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.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Qureshi MS, Sheikh QI, Hill R, Brown PE, Dickman MJ, Tzokov SB, Rice DW, Gjerde DT, Hornby DP
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