Figure 2

Preparation of MR-1 recombinant protein and anti-MR-1 polyclonal antibodies (PcAbs). (A) Construction of the of MR-1 expression plasmid. (B) The induced and purified MR-1 recombinant protein was visualized by 15% sodium dodecylsulfonate-polyacrylate gel electrophoresis (SDS-PAGE). Lanes 1-3: expressed products of MR-1 recombinant protein at 0 h, 2 h and 4 h; Lane 1': unpurified MR-1 protein; Lane 2': purified MR-1 protein; M: protein molecular weight ladder (170, 130, 100, 70, 55, 40, 35, 25, 15, 10 kDa). (C) The titer of anti-MR-1 PcAbs was tested by ELISA (R1: > 1:40,000; R2: > 1:80,000). (D) Western blotting of the MR-1 protein. Upper panel: MR-1 recombinant protein at 0 μg (a, d), 0.1 μg (b, e), 1 μg (c, f) was detected by anti-MR-1 PcAbs R1 and R2 at a dilution of 1:2000. Lower panel: Total protein from SKOV3 cell lysates at 0 μg (a'), 1 μg (b') and 10 μg (c') was detected by anti-MR-1 PcAb R2 at a dilution of 1:2000.