Fractionation and partial characterization of proteins extracted from the bovine fallopian tube: preparation of tools for further purifications

Described in the present paper is a combined biochemical and immunological approach to
study oviductal proteins in the bovine. Antisera were raised against semi-purified proteins extracted from bovine tubal mucosal tissue and were characterized. These antisera are available to monitor puritications of specific oviductal proteins in the future. Oviducts from 170 cyclic cows were collected at a slaughterhouse, and high amounts of mucosal proteins were extracted. The proteins were fractionated after precipitation with ammonium sulfate, anti-bovine serum albumin (bSA) and anti-bovine immunoglobulins bIg) afiinity chromatography and ion exchange chromatography. Each of the 12 fractions obtained after ion exchange chromatography was used to immunize a rabbit. Conditioned media were recovered from bovine oviduct cell monolayers cultured without serum to cot&m the oviductal origin of the extracted proteins. After Western blot analysis, 15 proteins were detected in the bovine oviductal extracts, and their molecular weights and isoelectric points were determined by 2 dimensional electrophoresis. Among these 15 proteins, 11 were also detected in conditioned media of bovine oviductal cells. These results demonstrate an oviductal origin of the 11 detected proteins and strongly suggest their secretion by the oviductal cells.