Production and purification of recombinant hFAM76B-6His.

(A) His6-tagged hFAM76B protein was produced in E. coli BL21 (DE3), purified by Ni–NTA column and subjected to 10% SDS-PAGE. Coomassie blue staining revealed a band of 42 kDa, consistent with the predicted size of hFAM76B. Lane 1, protein molecular weight marker; lane 2, nonbound sample; lanes 3–5, samples eluted with 5 mM, 50 mM and 250 mM imidazole, respectively. (B) The purified protein (50 ng/lane) was confirmed by Western blot analysis with anti-6His antibody and rabbit polyclonal anti-hFAM76B antibody (Sigma, St. Louis, MO).