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. Author manuscript; available in PMC: 2008 Dec 18.
Published in final edited form as: Biochemistry. 2007 Nov 17;46(50):14450–14460. doi: 10.1021/bi701310x

Fig. 1. SDS-PAGE of rFXI/C362S,C482S.

Fig. 1

Plasma FXI (150 μg) or rFXI/C362S,C482S (150 μg) was incubated with FXIIa (~7.5 μg) for ~16 hrs at 37°C and fractionated on SDS-PAGE (4–15% Tris-HCl) under reducing conditions and stained with Coomassie Blue. Lane 1, size markers (sizes denoted in the figure). Lane 2, pFXI and lane 3, pFXIa. The upper band is unactivated FXI, the middle band is the 50kDa heavy chain, and the lower band is the 30kDa catalytic domain. Lane 4 is a sample of the rFXIa/C362S,C482S mutant after activation by FXIIa (FXIIa was removed by passing the activation mixture over a corn trypsin inhibitor column). Lane 5, the heavy chain and unactivated rFXI/C362S,C482S eluted from a 5F7 antibody column. Lane 6 is the isolated catalytic domain after purification with the 5F7 column.