Dot blot with PVDF

Aawara Chowdhury via methods%40net.bio.net (by aawara from pontiff-playground.org)
Fri Feb 26 12:43:29 EST 2010


In <mailman.196.1267199236.25217.methods from net.bio.net>,
 Arne K Christensen <arnec from bio.umass.edu> wrote:

> I have run dot blots with nitrocellulose membrane before; simply add 
> 10ul samples in RSB directly to membrane and wait for them to dry, then 
> block and continue with the Western blot.  However, we only have PVDF 
> membranes on hand right now.  Has anybody here run dot blots on PVDF 
> membrane.  Do you apply the sample to the dry PVDF membrane before the 
> methanol soak?  Will proteins bind to the membrane before the methanol soak?

I have methanol soaked, then soaked in transfer buffer, than 
soaked in TTBS, and used it wet in the dot blotter with very very low
vacuum.

AC
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