Fig. 3

Purification and western blot analysis of BuLIF. A Purified recombinant BuLIF/His6x obtained from Lemo 21 (DE3) was analyzed using SDS-PAGE. After IPTG induction, cells were incubated for 12 h and harvested by centrifugation. Supernatant was collected after cell lysis and loaded on to the Ni-NTA resin. Approximately 23 kDa recombinant BuLIF/His6x was purified by affinity chromatography. Lanes 1 and 2 indicate protein marker and purified protein respectively. B Silver staining was done to analyze the purity of recombinant protein. A single band was observed after staining. C Purified protein was confirmed by antiLIF antibody in dilution of 1:1000. The band was developed using DAB (3,3′Diaminobenzidine) substrate on PDVF membrane after electro blotting. Lanes 1 and 2 indicate protein marker and confirmed recombinant protein respectively