Western Blot Smearing?

Jen jrobertson1 at sympatico.ca
Thu Aug 29 19:14:46 EST 2002


in my experience it is the incomplete dissolution of the milk powder
that leaves the little dark spots on my blots, so the longer i stir it
the less spots i get - you can try filtering your milk/tbs solution
through some regular old filter paper

however i prefer to use bsa :)


On Wed, 28 Aug 2002 07:26:11 +0100, Michael Witty
<mw132 at mole.bio.cam.ac.uk> wrote:

>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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