Many transcripts for proteins specifically expressed in haploid germ cells are synthesized in the spermatogenesis. A monoclonal antibody TRA54 recognizes specific organellaes in germ cell cytoplasm from spermatocytes to spermatids (1, 2). Western blots using TRA54 revealed broad bands with 85 kD, 190 kD and >200 kDa in testis (fig WB). This antibody can be used for immunohistochemical staining of testicular tissues(1, 3).
The hybridoma producing TRA54 was obtained from the immunaization of a rat with cell lysates of adult mouse testes by Prof. Yoshitake Nishimune group of Osaka University. This product is purified from the ascitic fluid of nude mice carrying the hybridoma clone TRA54.
1) Western blotting
2) Immunohistochemical staining
Analysis of various mouse tissues with TRA54 antibody. The antibody was 5,000-fold diluted before use.
Immunohistochemical staining of mouse testicular cross-section with TRA54 antibody.
Testis from C57BL/6 adult mouse was fixed in Bouin’s solution, followed by paraffin embedding and sectioning. After deparaffinized, sections were treated with normal goat serum, reacted with TRA54 antibody and incubated with biotinylated sheep anti-rat Ig antibody and biotin peroxydase/avidin, followed by incubation with hydrogen peroxide and diaminobenzidine.