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. Author manuscript; available in PMC: 2011 Apr 1.
Published in final edited form as: Nat Struct Mol Biol. 2010 Aug 22;17(10):1260–1262. doi: 10.1038/nsmb.1904

Figure 1. Purification of human BRCA2 and interaction with RAD51.

Figure 1

(a) Schematic representation of purified human BRCA2 protein. (b) BRCA2 purification analysis. Left: Silver-stained fractions from purification steps: I, lysate. II, pooled fraction after ammonium sulfate precipitation. III, pooled fraction of GSTrap eluate. IV, fraction from anti-FLAG column eluate. Right: Analysis of FLAG column load (L), flow through (FT), and fractions by PAGE and silver staining. (c) Immunoblotting with antibodies spanning the full-length tagged BRCA2 protein as indicated in a. (d) GST pull-down assay design. (e) Immunoblots and (f) quantitation of the proteins in pull-down assay. The reactions contain 4.65 nM (50 ng) or 9.30 nM (100 ng) RAD51, 0.1 nM (10 ng) or no BRCA2, and 1 mM AMP-PNP, ADP, or ATP. R.S.: Regenerating system. Error bars represent s.d. of n=3.