Figure 2.
Purification of RK from mammalian cells. (I) The protein extract from 5 × 108 transfected COS-1 cells was chromatographed on a heparin–sepharose column as in Materials and Methods. (II) Fractions 78–84 (14 ml) from I were diluted and applied to a Mono Q column. (III) Fractions 63–66 (8 ml) from II were diluted and applied to a Mono S column. (Inset) SDS/PAGE of selected fractions followed by protein detection by using Coomassie blue staining (A) or immunodetection (B) by using an anti-RK monoclonal antibody. L, preloading sample; M, molecular weight marker.