Figure 1
Characterization of recombinant PLAC1 and anti-PLAC1 antibodies. rhPLAC1was produced in E. coli and purified. The purity of the protein was assessed by SDS-PAGE analysis and Western blot using anti-His tag antibody (a) (full length gel and blot in supplementary information file, Fig. S6a,b). Monoclonal anti-PLAC1 antibody was generated in mice using rhPLAC1 as immunogen and reactivity of the purified anti-PLAC1 mAb, 2H12C12, was confirmed by indirect ELISA using rhPLAC1 as coating layer. Data were generated from at least three independent expeiments (b). Anti-PLAC1 mAb failed to react with PLAC1 in Western blotting and hence rabbit polyclonal anti-PLAC1 antibody was produced by immunizing the animals against rhPLAC1 and its reactivity with human placental PLAC1 was confirmed by Western blotting. In all lanes, lysate of human placenta has been loaded. Lanes 1, 2: Probed with purified rabbit anti-PLAC1 antibody. Lanes 3 and 4: Probed with PLAC1 hyperimmune rabbit serum. Lane 5: Probed with anti-PLAC1 mAb, 2H12C12. Lane 6: Probed with pre-immune rabbit IgG. Beta actin was used as loading control (c). After SDS-PAGE and transfer of proteins to the membrane, it was cut and different lanes were probed separately with different antibodies.
