[Arabidopsis] Question about CoIP

Wenxian Sun via arab-gen%40net.bio.net (by wenxian16 from yahoo.com)
Wed Feb 6 11:24:27 EST 2008


Hi Colleagues,

I am trying to use  CoIP to study the  protein-protein interaction in Arabidopsis seedlings. The proteins are tagged by HA and cMyc-GFP, respectively. When I use anti-cMyc antibody to do immunoprecipitation, I can detect the HA-tagged protein in the pulldown pretty well. However, if I use anti-HA antibody to do IP, I can not detect any cMyc-GFP tagged protein with anti-cMyc or GFP antibodies. The results are highly repeatable. The anti-GFP antibody is from Invitrogen. The anti-HA, cMyc antibodies are from Covance. I have tried protein A (Amersham), protein A/G (Pierce) beads and HA.11 affinity matrix (Covance) and have no luck. Anyone has the similar problem or any suggestion? 

Thanks a lot,

Wenxian
 
       
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