PMID: 2484046Oct 1, 1989Paper

Antibody against avian müllerian inhibiting substance (MIS) recognizes MIS on rat müllerian duct and human tumor cells

Proceedings of the National Science Council, Republic of China. Part B, Life Sciences
J J Wang, C S Teng

Abstract

Müllerian inhibiting substance (MIS) on rat Müllerian duct (Md), and four Müllerian-derived tumor cells: HeLa S-3, RL-95.2, A-431 and NIH:OVCAR-3 are recognized by the poly- and mono-clonal avian-MIS-antibodies (A-MIS-Abs) using avidin-biotin complex (ABC) immunolabeling techniques. Internalization of MIS-ligand complexes was successfully detected in HeLa S-3, OVCAR-3 and RL 95.2 cells. Control groups include: (i) the samples omitted primary antibody treatment, (ii) Wolffian duct by side of Md in the genital ridge, and (iii) another two MIS-negative tumor cell lines of non-Müllerian origin: Chang hepatoma ascites cell and mouse myeloma cell (X63-Ag 8.653). Genital ridges from rat embryos of 14d of gestation were removed under dissection microscope, fixed in 2.5% glutaraldehyde in D-PBS of pH 7.2 for 30 min, and sliced into 0.1-0.2 mm thick pieces. A-431, HeLa S-3 and NIH:OVCAR-3 were maintained in OPTI-MEM culture medium, RL-95.2 was cultured in F12 culture medium. The cells were transferred to 24-well flat bottom culture plates with Thermanox tissue culture coverslips. The immunolabeling of fixed and non-fixed samples were processed within the wells. These studies provide first immunocytochemical evidences for the similarity b...Continue Reading

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