Figure 1: SDS–PAGE analysis of IMAC purified recombinant Yersinia Outer Proteins (YOPs) and immunoreactivity with Y. pestis infected mice sera. [A] The panel depicts Coomassie stained recombinant purified YpkA (lane 2); YopH (lane 3); YopM (lane-4); V antigen (lane 5) and Pla (lane 6). Protein molecular weight markers were run in lane 1. [B] Immunoblot analysis of IgG antibody response to Y. pestis recombinant antigens. The purified recombinant antigens (2 μg each) were separated on 10% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was incubated with pooled sera (8 mice) at 1:1500 dilution with experimentally Y. pestis infected mice. The panel depicts YpkA (lane 2); YopH (lane 3); YopM (lane-4); V antigen (lane 5) and Pla (lane 6). Protein molecular weight markers were run in lane 1. [C] Immunoblot analysis of recombinant antigens with pre-infected mice sera at 1:1500 dilution. The panel depicts molecular weight markers (lane 1); YpkA (lane 2); YopH (lane 3); YopM (lane-4); V antigen (lane 5) and Pla (lane 6). [D] 12% SDS-PAGE analysis of recombinant YOPs. The panel depicts Coomassie stained recombinant purified YopN (lane 2); YopJ (lane 3); YopE (lane-4); YopK (lane 5) F1 (lane 6) and pH6 antigen (lane 7). Protein molecular weight markers were run in lane 1. [E] Immunoblot analysis of IgG antibody response to Y. pestis recombinant antigens. The purified recombinant antigens (2 μg each) were separated on 12% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was incubated with pooled sera (8 mice) at 1:1500 dilution with experimentally Y. pestis infected mice. The panel depicts YopN (lane 2); YopJ (lane 3); YopE (lane-4); YopK (lane 5) F1 (lane 6) and pH6 antigen (lane 7). Protein molecular weight markers were run in lane 1. Their sizes (in kDa) are shown at the left of the panel. [F] Immunoblot analysis of recombinant antigens with pre-infected mice sera at 1:1500 dilution. The panel depicts protein molecular weight markers (lane 1); YopN (lane 2); YopJ (lane 3); YopE (lane-4); YopK (lane 5) F1 (lane 6) and pH6 antigen (lane 7).