Purification and characterization of HLA(H)6 . (A) A 10% SDS gel stained with Coomassie Blue, on which aliquots of fractions containing HLA(H)6 were electrophoresed at different stages of its preparation. Lane 2, conditioned media from Cos cells transfected with pSG5-HLA(H)6 ; lane 3, flow-through from Ni-NTA-agarose column; lanes 4 and 5, wash fractions with and without Tween 20; lane 6, peak of eluate from Ni-NTA-agarose column; lane 7, eluate from Affi-Gel Blue; lane 8, flow-through from heparin-Sepharose. Lane 9 contains 1.0 μg of plasma-derived RSA, and lane 1 shows molecular weight markers (same as Fig 3). (B) Additional electrophoresis of aliquots of samples shown in lanes 7 and 9 in (A); the gel was cut in three, and portions stained with Coomassie Blue (Stain), immunoblotted with anti-RSA as in Fig 3, or blot-overlaid with thrombin (IIa).
