help with antibody

Graham Thomas gxt5 at PSU.EDU
Sat Nov 11 13:19:36 EST 1995


Artur Kania wrote

>.....Is there a way to modify the fixation to include a detergent or something
>which will denature my antigen ?  Should I use this detergent throughout
>the Ab staining protocol ? Has anyone encontered this problem before (and
>solved it) ?
>....

I believe that there is a mixed-micelles technique that I think may help in
this kind of situation. Add SDS to your detergent at 1/4-1/5th the
concentration of your triton - e.g. 0.02-0.033% SDS + 0.1% Triton.
Unfortunately I have no citation to give you; however, my notes indicate
that this method came originally (or at least to the person who told me)
from Larry Gerace at the Scripps so you might want to contact him if you
want to try this for more details (- and convey my sincere apologies if
this attribution is incorrect). I solved my problems in other ways and so I
cannot offer you the benefit of extensive experience in it's use.
        Maybe this will jog someones memory out there.
Good Luck
Graham

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