I am now using Sigma anti-flag M2 affinity gel to Pull-down my Flag-tagged protein from transgenic plant. I used 0.1M glycine HCl,pH3.5 to elute the binding protein as Sigma suggested. But I got huge amount of antibody bands (around 50kD and 25kD)and little or nothing for my tagged protein when I used anti-flag M2 antibody for Western blot. However, I got clear result when I used the same antibody to detect my protein in protein extract from transgenic plant. Does anybody have experiences with this affinity gel or any suggestions?
Thanks in advance.
Yajie Niu
Graduate Student
Washington State University
