Purification of "inhibin" from human ovarian follicular fluid.
Following the earlier demonstration of inhibin-like activity in human ovarian follicular fluid a method for its purification to apparent homogeneity is described. The fluid was converted to acetone powder and subjected sequentially to ammonium sulphate fractionation, gel chromatography on Sephadex G-200, continuous gradient ion-exchange chromatography on DEAE-cellulose, first with a pH gradient from 8.0 to 4.0 and then with a NaCl gradient to 1 M at pH 5.2. The active fraction from this step was subjected to gel filtration on Sephadex G-100 and finally passed through an Amicon Centriflo membrane CF-25 (cut off point: 25,000 m.w.). The ultrafiltrate was homogeneous by SDS-polyacrylamide gel electrophoresis, had a molecular weight of the order of 23 000 and was capable of suppressing serum gonadotrophin levels in the castrated male rats in as low a dose as 25 microgram/rat.
D000328:Adult; D000818:Animals; D001681:Biological Assay; D001826:Body Fluids / Q000032:analysis*; D002369:Castration; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004589:Electrophoresis, Disc; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005374:Filtration / Q000295:instrumentation; D005640:Follicle Stimulating Hormone / Q000097:blood; D006728:Hormones / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D008970:Molecular Weight; D006080:Ovarian Follicle / Q000032:analysis*; D011863:Radioimmunoassay; D051381:Rats; D013972:Thyrotropin / Q000097:blood
D006728:Hormones; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D013972:Thyrotropin