non-block with PVDF ??

Phil Terman gambler at cts.com
Mon Nov 13 21:29:06 EST 1995


Stefan Baeckstroem <seab> wrote:
>Hello there.
>
>Has anyone tested the hydrophobic non-block method with western when using PVDF
>membranes published in a recent number of analytical biochemistry (sponsored by
>millipore) ?
>
>They say that you can save time, but it seems like it takes longer time to me.
>After the blot you have to dry the membrane properly, which they recomend you
>to do in rt overnight. Then you have earned two hours with the non-block, but
>you lost one day having to wait for the membrane to dry before the
>immunodetection.
>
>Any comments ?
>--
>		Stefan Backstrom
>		seab at nucleus.ibg.uu.se
>		http://www.ibg.uu.se/~seab
>
*********************************************************************************************
That is one way in which the membrane can be dried but of course you already figured out you lose more time...
To ensure you save the time that you meant to in the first place you can 1)soak the membrane in 100%methanol
for 10 seconds to drive out the water, place blot on a piece of filter paper and then wait for the methanol to evaporate...This drying
method only takes 15 minutes.............Other way to dry would be to place the blot in a vacuum chamber (blot between
two sheets of filter paper)...this would take about 30 minutes...........The drying just reduces the background.
It does work for me but I prefer PVDF over crackly nitrocellulose anyday of the week however i use it...Good Luck!



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