Western Blot Smearing?

Michael Witty mw132 at mole.bio.cam.ac.uk
Wed Aug 28 01:26:11 EST 2002


On Wed, 28 Aug 2002, Jen wrote:

>
> sounds like a blocking problem
>
> are you using skim milk powder as a block? if so i usually have to
> stir it on high for at least an hour before blocking.
>
> i get much better results with 5% BSA in TBST + 0.02% Na-azide (so I
> can reuse it multiple times)

Dear Jen,
        I just add the milk powder to TBS, dissolve the milk powder
(sometimes not completely) and add the PVDF or nitrocellulose membrane.
Why do you stir for >1hour before adding to the membrane?  Regards, Mike.




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