identification and purification of hormone - protocols + knowledges needed about hormone (Jun/05/2006 )

Hi everyone,

I want to know if my cells secreted hormone in the culture medium. If so, I want to put a name on the hormone. Is anybody has ever worked on this kind of experience and has some protocols and paper to share with me?

Thanks! :-)

What I did was to try to immunoprecipitate the hormone from the media.. After filtering the media I incubated overnight at 4 with the 1st antibody and then for 4 hrs with protein A agarose. Centrifugue, discard supernatant, wash 7 times and resuspend pellet in loading buffer for SDS page. Sorry I dont have numbers and quantities but I adapted the protocol from the one given by ROCHE protein A agarose.
hope this helps a bit