Skip to main content
Figure 3 | BMC Immunology

Figure 3

From: Serum from mice immunized in the context of Treg inhibition identifies DEK as a neuroblastoma tumor antigen

Figure 3

SDS-PAGE and Western Blot Analysis of Recombinant DEK. (A) Bacterial lysates and Ni-NTA column purified protein were diluted in reducing sample loading buffer, resolved by SDS-PAGE and stained with Coomassie blue. Lanes were loaded with 1) molecular weight (mw) marker, 2) control bacterial lysate, 10 μl, 3) pET-15b/DEK lysate, 10 μl, 4) 1.5 μg Ni-NTA column-purified DEK, 5) pET-15b/EGFP lysate, 10 μl, 6) 1.5 μg Ni-NTA column-purified EGFP. (B) Proteins were resolved by SDS-PAGE and then transferred to PVDF membrane using a Bis-Tris electrophoresis buffer system for western blot analysis. In blot (a) lanes contained: 1) mw marker, 2) control bacterial lysate, 3) pET-15b/DEK lysate, 10 μl, 4) 0.15 μg Ni-NTA column-purified DEK, 5) pET-15b/EGFP lysate, 10 μl, 6) 0.15 μg Ni-NTA column-purified EGFP. Blot (a) was probed with anti-6x-His antibody. For blot (b) a separate gel containing the same samples loaded in blot (a) was probed with anti-human DEK monoclonal antibody. Bound antibody was detected in both blots with alkaline phosphatase (AP)-conjugated rabbit anti-mouse IgG (H+L) secondary antibody, as described in Methods. Data is representative of more than three separate experiments.

Back to article page