Weird Western Blot (circular streaks)

Paul S. Brookes. brookes at uab.edu
Thu Jul 20 12:03:49 EST 2000


Sounds like air bubbles in the transfer cassette.   Also if you're using 
PVDF make sure you soak it in MeOH first, as it doesn't wet easily in TX 
buffer.

It is also possible the fiber pads that were provided with the original set 
up are expired.  They get thin after a while so the pressure between 
gel/membrane is not even.  They also get caked up with proteins which can 
cause interference with certain western blots (for example 
nitrotyrosine).   You can solve this by boiling them for 1 hour in a 1 
liter beaker of de-ionized water.  They come out all white and fluffy, just 
like new, and considerably thicker than before so they'll apply more 
pressure to your blot assembly.   And at $20 for a new set of what appear 
to be 50 cent kitchen scouring pads, rejuvenating them this way saves you 
money too.

_________________________________________
Dr. Paul S. Brookes.            (brookes at uab.edu)
UAB Department of Pathology,   G004 Volker Hall
1670 University Blvd., Birmingham AL 35294 USA
Tel (001) 205 934 1915     Fax (001) 205 934 1775
http://peir.path.uab.edu/brookes

The quality of e-mails can go down as well as up
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