Figure 2

Expression of recombinant Ldccys2 using insect cell expression system. Western blots showing recombinant expression of Ldccys2 using anti-JHE antibody (A) and duplicate blot probed with anti-Ldccys2 antibody (B). The lanes are labeled appropriately. 5 μg of supernatant from the insect cells expressing recombinant plasmids or the control were separated on 8% SDS-PAGE, blotted onto to PVDF membranes and probed with 1:500 dilution of anti-JHE or 1:100 dilution of anti-Ldccys2 antibody. A horseradish peroxidase conjugated anti-mouse secondary antibody was used and the blot was developed by using ECL chemiluminescent kit. (C) Gelatin assay showing cysteine protease activity of the recombinant protein. The supernatant was separated on a gelatin gel and incubated in a reducing buffer. The gel was then stained with Coomassie Blue to visualize the clearing of gelatin on the gel indicating cysteine protease activity.