Fig. 3.

Characterization of 12 IgG-type monoclonal antibodies. A Isotypes of 12 mouse IgG-type monoclonal antibodies (IgG1, IgG2a, IgG2b, IgG3, and kappa/lambda) derived from hybridoma supernatant of the 12 clones. B Amino acid sequences of mouse immunoglobulin heavy and light chain variable regions of 12 mAbs. PCR fragments of heavy and light chain variable region genes were amplified from cDNA derived from hybridoma clones by using IgG variable region-based degenerate PCR primers as shown in Additional file 1: Table S1. The consensus sequences of IgG light chain (a) and IgG heavy chain (b) of variable regions were compared. Three complementarity determining regions in light and heavy chains (CDR-L1-3 and CDR-H1-3) were shown in shading boxes. C Identification of the antigenic molecules recognized by representative mAbs. Cell surface were biotinylated and antigenic molecules were captured by mAb. These molecules were visualized by western blot using streptavidin-HRP