Gary schrieb:
>> I'm trying to strip a nitrocellulose membrane [10 mM b-mercaptoethanol, 0.1%
> SDS, 20 mM Tris, pH 6.5] to restain it with another antibody . The signals
> I 'm looking for are phosphoproteins. I appear to be losing more signal than
> I would expect. Is there a chance that I'm hydrolyzing the phosphate group
> during stripping? How do I avoid this? Thanks.
>> Gary
Hi Gary !
I do also a lot stripping, and I´m using the same protocol as You
describe here. Although I´m not looking for Phosphoproteins, I
encountered the same problem that the signal strength is decreasing due
to stripping the Blot. Therefore I guess that it´s rather a general
problem of destroying the relevant epitopes the antibodoies recognizes
than a problem of hydrolyzing the Phosphates........
Hope this information helps You something.....
Markus
