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Updated: May 12, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells
Published on: December 9, 2016
Affinity filtration coupled with capillary-based affinity purification for the isolation of protein complexes
M S Qureshi1, Q I Sheikh, R Hill
1Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, UK.
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.
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