Stripping nitrocellulose membranes containing phosphoprotein

Meißner Markus meissner.markus at t-online.de
Sun May 14 14:27:14 EST 2000


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




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