Protocol

Adhesion Protein Protocols

Volume 1046 of the series Methods in Molecular Biology pp 231-250

Date:

Purification of Native Arp2/3 Complex from Bovine Thymus

  • Lynda K. DoolittleAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute
  • , Michael K. RosenAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute
  • , Shae B. PadrickAffiliated withDepartment of Biophysics, UT Southwestern Medical Center and Howard Hughes Medical Institute

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Abstract

The Arp2/3 complex is an actin filament nucleator involved in cell motility and vesicle trafficking. Owing to the role the complex plays in important and fundamental cell biological processes, the purified complex is used in biochemical assays, reconstituted motility assays, and structural biology. As this is a eukaryotic complex assembled from seven polypeptides, the complex is purified from eukaryotic sources. Described here is a detailed method for purification of the complex from a mammalian tissue, bovine thymus.

Key words

Actin nucleation factor Arp2/3 complex Biochemical purification Endogenous source Detailed protocol