Immunotherapy-induced antibodies to endogenous retroviral envelope glycoprotein confer tumor protection in mice
Fig 2
Modified 2D immunoblotting allows for the identification of tumor antigens.
(A) The schematic describes the workflow to partially transfer a 2D gel and overlay silver-stained gel on chromogenic immunoblot to identify the spots of interest. (B) B16F10 membrane proteins and biotinylated BSA, soybean trypsin inhibitor, and equine hemoglobin were immunoblotted with CM7 serum and detected with IRDye-800CW-conjugated anti-mouse IgG2b and IgG2c Abs (green) and Alexa-Fluor-647-conjugated streptavidin (red). (C) Identical sample in (B) was partially transferred and chromogenically detected with biotinylated anti-mouse IgG2b and IgG2c Abs and HRP-conjugated streptavidin. (D) The gel remaining after the partial transfer was silver-stained and overlaid on the chromogenic immunoblot. Red boxes indicate the spots of interest.