Western Blot with HPLC purified samples - non specific binding??? need help desperately (Jun/26/2008 )

Dear all, I need some help with Western Blot because I'm totally new to it. My problem was the results showed more bands than I expected and I'm not really sure if it is due to non-specific binding as these bands are rather dark in intensity. Below is the image of my results and wonder if anyone could tell me if those with red arrows pointing towards it are non-specific binding or are they due to binding with the primary antibody?

In addition, is it normal/correct to see my proteins on the gel after transfer (stained with Coomassie Blue)?



Hope I can get some replies.

Thanks in advance!

Dear all, I need some help with Western Blot because I'm totally new to it. My problem was the results showed more bands than I expected and I'm not really sure if it is due to non-specific binding as these bands are rather dark in intensity. Below is the image of my results and wonder if anyone could tell me if those with red arrows pointing towards it are non-specific binding or are they due to binding with the primary antibody?

In addition, is it normal/correct to see my proteins on the gel after transfer (stained with Coomassie Blue)?



Hope I can get some replies.

Thanks in advance!