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. 2004 Sep 16;101(39):14180–14185. doi: 10.1073/pnas.0405878101

Fig. 1.

Fig. 1.

His-tagged Btk is functional and can be purified with a Ni-column. (A) Btk– DT-40 cells were transduced with WT or 6× His-tagged Btk, and the calcium response was measured with anti-IgM (2.5 μg/ml) stimulation. (B)6× His-tagged Btk was coexpressed with activated Lyn (Y508F) in 293T cells and purified through a Ni-column. The purified fraction was separated on SDS/PAGE, stained with Coomassie blue, and confirmed by Western blot with anti-Btk antibody. (C) 10× His-tagged Btk was infected into Btk-deficient DT-40 cells, and purified Btk was stained with Coomassie blue and confirmed by Western blot. (D) Phosphopeptides identified by MS. P, phosphorylation.