title: Method for isolating pure bovine gonadotrophs from anterior pituitary using magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor antibody.
abstract: No methods are currently available for rapidly isolating gonadotrophs from the anterior pituitary (AP) in any species. We developed a method for preparing pure bovine gonadotrophs from a heterogeneous AP cell mixture by magnetic separation and our original antibody against the N terminus of bovine gonadotropin-releasing hormone receptor (GnRHR). A bovine AP cell mixture was incubated with the anti-GnRHR antibody, anti-dextran antibody-conjugated secondary antibody and dextran-coated magnetic nanoparticles for magnetic isolation. Approximately 5.2 × 106cells were isolated per AP of Japanese Black heifers (26 months of age) and cultured, and confocal microscopy confirmed to be GnRHR- and luteinizing hormone-positive, corresponding to a purity of 100%. Approximately 44.5 µg of total protein was extracted from the pure gonadotrophs per AP.
authors.affiliation: Joint Faculty of Veterinary Medicine, Yamaguchi University, Yoshida 1677-1, Yamaguchi-shi, Yamaguchi 753-8515, Japan.
authors.forename: Kiran
authors.initials: K
authors.lastname: Pandey
authors.forename: Ashrafun
authors.initials: A
authors.lastname: Nahar
authors.forename: Hiroya
authors.initials: H
authors.lastname: Kadokawa
mesh_terms: D000818:Animals; D000906:Antibodies; D002417:Cattle; D002478:Cells, Cultured; D005260:Female; D052681:Gonadotrophs / Q000166:cytology*; D018189:Immunomagnetic Separation / Q000379:methods; D007986:Luteinizing Hormone / Q000378:metabolism; D058185:Magnetite Nanoparticles; D010903:Pituitary Gland, Anterior / Q000166:cytology*; D011966:Receptors, LHRH / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D058185:Magnetite Nanoparticles; D011966:Receptors, LHRH; D007986:Luteinizing Hormone