SDS-PAGE ghost band help - (Mar/29/2006 )
I would like some help as to what could be causing ghost bands. I am running an SDS-PAGE 4-20% tris-gly gel with tris-gly buffer, I have a blank lane between my markers and my sample but are picking up distinct bands in the blank lane that correspond to the low mw bands in the sample. the load volume is very small, 4microliters. the bands are visualized by coomassie staining but when i do a western blot, they are not detected, are they real?
It might be transfer from the sample lane to the blank lane. This can happen if you have bubbles in your solution when you load (and can also occur if your pipette tip is not properly inside the well). It's quite an easy thing to do actually.
I guess they're not in the Western because they either don't react with the antibody or there simply isn't enough (which wouldn't surprise me).