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. 2018 Jul 18;9(8):791. doi: 10.1038/s41419-018-0824-2

Fig. 1. Identification of BAP31 as a new CTA using the spermatogenic cell-specific monoclonal antibody-defined cancer/testis antigen (SADA) method.

Fig. 1

The SADA method was employed to generate candidate antibodies against cancer/testis antigens. Spermatogenic cells purified from donated testis tissues were used to immunize BALB/c mice, and hybridomics procedures were used to generate candidate antibodies against Spermatogenic cells. a The antigen recognized by FM-1 was confirmed as a CTA by immunohistochemical staining of testicular tissues, somatic normal tissues, and related cancer tissues. The scale bars represent 100 μm. b SDS-PAGE electrophoresis and silver staining of the FM-1 immunoprecipitates from HeLa cells lysates revealed that an ~ 28 kDa protein band (shown by arrow) was enriched in the gel. c Immunoprecipitates from HeLa cells lysates treated with FM-1 were analyzed with an FM-1 antibody and anti-BAP31 antibody by western blots. d CHO cells were transfected with the BAP31-GST plasmid pcDNA3.1. Cell lysates were subjected to western blotting with FM-1, anti-GST, and anti-BAP31 antibodies, respectively. HeLa cells lysates were loaded as a positive control