Protocol Online logo
Top : New Forum Archives (2009-): : Protein and Proteomics

coomassie blue stain - (Jul/10/2013 )

hello everybody,

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?

thank you


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.