Gel electrophoresis and Western blotting of GST-fused b5R and GST-fused Cys203Tyr b5R.

(A) Gel electrophoresis of GST-fused b5Rs expressed in E. coliBL21. A total of 5 μg of GST-fused b5R or GST-fused Cys203Tyr b5R purified with glutathione Sepharose 4B was subjected to electrophoresis on 10% SDS-polyacrylamide gel. Lane 1, molecular weight markers used are phosphorylase B (94 kd), bovine serum albumin (67 kd), actin (43 kd), and bovine carbonic anhydrase (31 kd). Lane 2, GST-fused Cys203Tyr b5R. Lane 3, GST-fused wild-type b5R. (B) Western blotting of GST-fused b5R and GST-fused Cys203Tyr b5R purified with glutathione Sepharose 4B. Lane 1, GST-fused wild-type b5R. Lane 2, known rb5R as control (see text). Lane 3, GST-fused Cys203Tyr b5R.