Using Vitellogenin polyclonal antibody ( Cat # PAB13582 ) at a 1 : 4,000 dilution. ( 1 ) Plasma from male sole ( load 0.11µl of plasma ).( 2 ) Plasma from vitellogenic female sole ( load 0.11µl of plasma ).( 3 ) Plasma from estradiol-treated male sole ( load 0.08µl of plasma ).( 4 ) Vitellogenin precipitate ( load 0.83µl of precipitate ).( 5 ) Purified Vitellogenin ( load 2µl of purified preparation ).