Molecular diversity of TEX101, a marker glycoprotein for germ cells monitored with monoclonal antibodies: variety of the molecular characteristics according to subcellular localization within the mouse testis.

摘要: Abstract TEX101 was characterized as a unique germ cell marker molecule using the specific monoclonal antibody (mAb), TES101. Although this mAb has strong affinity/specificity for TEX101, TES101 loses its reactivity under reducing conditions. In study, we have generated new mAbs against to compensate shortcomings of different approaches. First, immunized mice with antigen on baculovirus expression system and isolated anti-TEX101 mAbs, 6002 6035. Second, raised Ts4 from spleen cells an immunologically naive old mouse. Western blot analysis revealed that possess immunoreactivity reducing/non-reducing Immunopositive staining Bouin-fixed sections observed in spermatocytes, spermatids testicular spermatozoa, but not other cells, similar paraformaldehyde (PFA)-fixed frozen stained previously reported. However, whereas 6002/6035 mainly showed only spermatocytes PFA-fixed sections, spermatozoa clearly recovered when were autoclaved or treated SDS. Peptide mapping deglycosylation indicated epitopes TES101, 6035 are located within 25–94 , recognized N -linked carbohydrate moieties Triton X-100-soluble mouse extracts extracellular water-soluble fractions. These results suggest strongly molecular association structure varies according subcellular localization seminiferous tubules. will be valuable tools further including function(s).