title: A reference preparation of buffalo pituitary follicle stimulating hormone using lectin affinity chromatography.
abstract: An improved and cost effective method to isolate FSH from buffalo pituitary glands is described here. The buFSH activity was monitored throughout by a highly sensitive heterologous radioimmunoassay (sensitivity 0.2 ng oFSH/mL) and the in vivo biological activity of the final preparation was also established. A biologically active buFSH-enriched preparation with a moderate recovery (42%) was obtained. The yield of the final buFSH-enriched preparation was 26.5 mg/kg of buffalo pituitary gland. In SDS-PAGE, the purified buFSH resolved as a heterodimer of 30 kDa molecular size, with a 21 kDa presumptive alpha-subunit. This preparation was also characterized in terms of biological and some of its physicochemical properties. A high-titer antiserum to buFSH was also raised in rabbit using this preparation. The reagents generated, buFSH and buFSH-specific polyclonal antisera, have possible diagnostic and therapeutic usage for improvement of reproductive health of water buffaloes.
authors.affiliation: Hormone Research Laboratory, Department of Zoology, University of Delhi, Delhi, India.
authors.forename: Hitendra S
authors.initials: HS
authors.lastname: Chand
authors.forename: Rajesh
authors.initials: R
authors.lastname: Chaudhary
authors.forename: K
authors.initials: K
authors.lastname: Muralidhar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D002020:Buffaloes; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D005260:Female; D005640:Follicle Stimulating Hormone / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D009929:Organ Size / Q000187:drug effects; D010053:Ovary / Q000187:drug effects / Q000254:growth & development; D010902:Pituitary Gland / Q000737:chemistry*; D011817:Rabbits; D011863:Radioimmunoassay; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives; D014611:Vaccination
chemical_list: D000911:Antibodies, Monoclonal; D007106:Immune Sera; C028326:concanavalin A-sepharose; D005640:Follicle Stimulating Hormone; D012685:Sepharose