> > Such high salt might neutralize static charges on IgG, but why would
this
> help in binding epitope. Surely, the closer to physiological conditions
the
> better. Am I missing somthing? The person who designed this buffer must
have
> had a good reason and I don't want to just ditch a protocol without a
clear
> understanding of the whys and wherefores.
Sometimes protocols are just hand-me-downs of 'what has worked before'. If
you have a new one that works - great, but if something goes wrong then it's
you and only you who can figure out what went wrong, cause no one else has
done this before.
As to salt and higher affinity - it could be that your AB-epitope
interactions are mostly hydrophobic, which means that they will be enhanced
(greatly) by higher salt, for obvious reasons.
A.G.E.
