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  Indian J Med Microbiol
 

Figure 1: (a) Confirmation of KFD NS1 protein expression by Coomassie staining of 12.5% SDS PAGE. Lane 1: Lysate of uninduced BL21 (DE3) cells; Lane 2: KFD NS1 protein expressed in BL21 (DE3) cells after induction with 1mM IPTG at 37°C for 4 h; Lanes 3, 4, 5: Elution fractions 1, 2 and 3 of induced cell lysate purified using Nickle chelating column; and Lane 6: Molecular weight marker; (b): Confirmation of KFD NS1 protein expression by Western Blot using commercially available anti-histidine monoclonal antibody. Lane 1: Protein lysate before purification showing reactivity with anti-histidine antibody; Lane 2: KFD NS1 protein showing reactivity at expected size (42 kDa) with anti-histidine monoclonal antibody; and (c) Confirmation of KFD NS1 protein expression by Western Blot using human serum sample positive for anti-KFD IgM and IgG antibodies, thereby increasing the probability of the presence of anti-KFD NS1 antibodies. Lane 1: Protein lysate before purification showing reactivity with KFD positive human serum; Lane 2: KFD NS1 protein showing reactivity at expected size (42kDa) with KFD positive human serum.

Figure 1: (a) Confirmation of KFD NS1 protein expression by Coomassie staining of 12.5% SDS PAGE. Lane 1: Lysate of uninduced BL21 (DE3) cells; Lane 2: KFD NS1 protein expressed in BL21 (DE3) cells after induction with 1mM IPTG at 37°C for 4 h; Lanes 3, 4, 5: Elution fractions 1, 2 and 3 of induced cell lysate purified using Nickle chelating column; and Lane 6: Molecular weight marker; (b): Confirmation of KFD NS1 protein expression by Western Blot using commercially available anti-histidine monoclonal antibody. Lane 1: Protein lysate before purification showing reactivity with anti-histidine antibody; Lane 2: KFD NS1 protein showing reactivity at expected size (42 kDa) with anti-histidine monoclonal antibody; and (c) Confirmation of KFD NS1 protein expression by Western Blot using human serum sample positive for anti-KFD IgM and IgG antibodies, thereby increasing the probability of the presence of anti-KFD NS1 antibodies. Lane 1: Protein lysate before purification showing reactivity with KFD positive human serum; Lane 2: KFD NS1 protein showing reactivity at expected size (42kDa) with KFD positive human serum.