I did a Immunoprecipitation then follow by western lot. When I use antibody A, I can detect all the inputs except 2 (HA-tag and truncated, thus antibody A cannot detect them). I can also see the interaction when I pull down with M2 beads (same case, when I probe with antibody A, I can see the interaction except the 2 that has HA-tag and truncated.
So, I went on to use HA antibody to probe for all the inputs since everyone has HA tag. I use 1:200 primary HA antibody and I can see all the inputs. However, I couldnt detect the interaction like I previously probe with antibody A on the M2 pulldown.
What might tbe the problems?
Thanks for help guys
Western blot antibody
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