Fig. 1
Cloning, expression, immunoreactivity, and cytotoxicity of rMuIFNλ3 protein expressed by E. coli. Colony PCR of the transformed E. coli BL21 (DE3) cells harboring pHis-MuIFNλ3 plasmid showing 582 bp product corresponding to the size of MuIFNλ3 gene (a). SDS PAGE analysis of IPTG-induced protein expression in the cell lysate of E. coli cells showing (closed box) the presence of rMuIFNλ3 protein with a molecular weight of ~ 25 kDa (b). Different elution fractions of Ni–NTA affinity column purified rMuIFNλ3 protein (E1 to E9, closed box) (c). Western blot analysis of purified His-tagged rMuIFNλ3 probed with mouse monoclonal anti-His antibody confirms the identity and size of the MuIFNλ3 protein (d). Schematic of the rabbit immunization schedule to raise hyperimmune sera against rMuIFNλ3 protein (e). Western blot analysis shows immunoreactivity of rMuIFNλ3 protein probed with rabbit hyperimmune sera. (f). Determination of CC50 of rMuIFNλ3 protein in murine B16F10 (65 μg/mL) (g). Each point represents the mean of three independent data sets ± standard deviation (SD) of the means