Fig. 3

Immunoprecipitation and Ig class determination of MS clones. a Immunoprecipitation was performed using 15 representative clones obtained by MS combination, and the precipitants were subjected to SDS-PAGE followed by WB with HRP-anti-mouse IgG antibody. The samples indicated by arrows are antibodies that did not give signals in WB; however, all mAbs, including these, were antibodies that could be used for immunoprecipitation. For comparison, the results of immunoprecipitation using antibodies obtained by EW are shown; however, only one of the three mAbs could be used for immunoprecipitation. b Ig class determination. After electrophoresis of the culture supernatant of the hybridomas of the MS clone by SDS-PAGE, WB was performed with HRP anti-mouse Ig antibody. The results showed that all mAbs used in the immunoprecipitation test were from the IgG class, as they all showed bands of H chain of approximately 55 kD and L chain of approximately 25 kD. The Ig class results for the other MS clones are shown in the additional file. The mAbs of the clones obtained by EW were also from the IgG class