coomassie blue stain - (Jul/10/2013 )
i did co-immunoprecipitation using agarose A beads and wants to verify my IP by coomassie blue staining. My questions are:
1. I used agarose A beads but my antibody is monocolonal, will this affect my result?
2. I performed western blot last time and found that my target protein co-immunoprecipitated with my IP antibody. I repeat the experiment again and this time wants to stain it with coomassie blue. I only found one band in my IP lanes and it seems to correspond to my target protein (~55Kda).
my question is, could this be my target protein or will the IgG band also show in the coomassie blue stain?
1. no, it shouldn't make a significant difference
2. the igg should show if there is enough to be detected with coomassie. you can silver stain to confirm the presence of the igg.