FIG. 4.

Generation of anti-PCARDv1 MAbs. (A) The FLISA (in-cell Western) results of PCARDv-1 on transfected Cos-7 cells with sera of mice immunized with plasmid harboring PCARDv1 and control cells (Irr. transfected), cells transfected with irrelevant plasmid Mns1 (meiosis-specific nuclear structural encoding pcDNA plasmid). No Pri, no primary antibodies added, only secondary antibodies added; Pre Im, the pooled sera of mice collected prior immunizations; m1–m5, sera from 5 different immunized mice. The p values of titers of m1, m2, m3, and m5 when compared to cells transfected with irrelevant plasmid were <0.01. (B) FLISA image of transfected Cos-7 cells expressing PCARDv1 plated in one representative 96-well plate showing positive clones. Undiluted supernatants from harvested hybridoma cell lines were used to screen for clones producing anti-PCARDv-1 antibodies. (C) Western blot image of anti-PCARDv1 antibody activity against various cell and tissue samples expressing PCARDv1. Fifteen positive clones resulted in this screen.