stripping and western blot?

Emir Khatipov khatipovNO at NOuchicago.edu
Fri Aug 23 16:08:17 EST 2002


1) I don't incubate at 50C for more than 15 min. It might be that you are
ovestripping the membrane, especially if your initial signal was low.
2) Sometimes BME stripping does not work. Try stripping in 2M NaOH for 30
min at R/T.
Emir

"leezuyoug" <leezuyoug at yahoo.com> wrote in message
news:12434ec2.0208230543.243c2b77 at posting.google.com...
> Hi, I have met some problems in my stripping of PVDF membrane. I use
> the protocol as follows:  50ml 62.5mM Tris-HCl (62.5), 2% SDS, 100mM
> 2-mecaptoethanol under 50C for 30 minutes. After stripping, wash 6x 5
> minutes, then reprob again. But it is always blank without any
> signal!!!
>     I am working at p38 which is one kind of MAPK, using the ECL
> reagent system.  Is there anyone can help me about this?
>      thinks a lot!!!





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