Immunoprobing Question / Data Interpretation - (Apr/08/2007 )
I have been doing western blots on serum samples that represent different timepoints after drug treatment. I have noticed that three of my samples often give a band around the same molecular weight for different antibodies. At first I thought there was something happening during those timepoints but now I am beginning to wonder if these bands are real? I know that these bands are not IgG. Is it possible that the proteins I am probing for share a similar sequence that is recognized by many antibodies? Any advice or thoughts in what may be happening is greatly appreciated!
it could be shared epitopes (or fragments of the protein) or non-specific binding.
if it is non-specific binding then you may remedy it by using more dilute primary and/or increasing the tween in your dilution buffer.
How do you know they aren't IgG?
Of course it's possible the regions could be similiar..just run a BLAST for proteins if you know the sequence. Of course you haven't told us much..so it could be anything really..sorry