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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