Diffusion of Antibodies in PVDF and NZ membranes

[Kyle Legate]

WS wrote:
>
> I'd recommend changing (i.e. normally increasing) the salt
> concentration in the buffers. Adding ethanol or methanol to the buffer
> (as you probably do it during the electroblot) also might help, but it
> could denature your antibodies and / or kill your detection system. It
> must be a pain to find out (how does one do that???) that your
> favorite protein does not like the standard conditions. 
> 
I'd use a dot blot approach to screen conditions. I never thought of 
that, silly me. I have a homemade antibody that gives a single strong 
cross-reactive band in brain lysate and a lot of minor background 
binders in cell lysate. Maybe I'll try the above approach this weekend 
to try to improve the specificity. I was resigned to the probability 
that I'd have to try and raise it again in a new rabbit--maybe I won't 
have to.