Antibody Mismatch Tolerance - (Apr/06/2007 )
I am using an antibody which has been raised against a synthetic peptide matching the human sequence, but am also working in mouse. There is one mismatch and two conservative substitutions in the mouse at the target sequence. Is the antibody still likely to work? What is the tolerance of an antibody to sequence differences?
This will depend on the effect of mismatch and substitution in the correct folding and return of an exact target epitope. Therefore the tolarance depends on the effect of substitution and the intial role of the replace amino acid.
It's the classic "how long is a piece of string?" question. You will just have to try it and see. If things don't go well, you could try changing the wash conditions, but you may get more non-specific binding.