liquid or semidry western blotting?

Paul S. Brookes. brookes at uab.edu
Thu Oct 5 17:42:58 EST 2000


As usual, you get what you pay for - $750 for a semi dry system from BioRad 
vs. $375 for the wet type apparatus, or even cheaper if you already have 
the mini-tank and only need the "guts".

If you transferred through the PVDF, its probably a function of the PVDF 
(maybe overloaded).  I can't see what all the fuss is about with PVDF - 
we've used NC for everything and never had any problems, then we switched 
to PVDF and got into all sorts of trouble with things not sticking to it, 
so we switched back after a month.  NC is cheaper too.   Voltage wise, if 
you include a bit of SDS in the buffer then most proteins (200kD or below) 
will go at 100V for 1.5hrs or 40V overnight.

For something inert to take up the slack volume, try a chunk of polystyrene 
culled from an ice bucket and sealed insie a zip-loc bag, or you could even 
use a zip-loc bag full of ice, thereby improving the cooling.

PSB

_________________________________________
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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