HELP! This is my first attempt to immunoprecipitate insulin receptor beta using protein A sepharose. My initial attempts using the IP kit from Upstate have all failed. I ordered Protein A Sepharose from Amersham but now I dont know what to do with it. Does anyone have a detailed protocol on how to use it? It is in a solution of ethanol, but I assume this has to be washed. And what does it mean to "preclear" my lysate?
Any help would be greatly appreciated!!