Western after Ip- no IP protein, but .... - (Jun/05/2006 )
I am doing IP with anti-Src Ab, then I perform a western to reveal some complex formation (like, ERK,et cetera), so I do see the coimunoprecipitated proteins but NO Src itself???!!!
If it can be that Src is not dissociates from the Ab in sample buffer+boiling???????
I cannot believe it is not dissociated.
2 possibilities :
-either your ant-src is not good. do you load a total extract to have a positive control?
-either your IP is not good, and what you see is not co-IP, but some proteins that bind to the agarose beads, did you add a control of IP without antibody?