NEPHGE and SDS transfer

[Stephen P. Driska PhD]

rep at icbr.ifas.ufl.edu wrote:

: Hey there,
: 	Happy Holidays and all that.  Posting this question for a friend.  Has
: anybody done NEPHGE gels and then transferred them under SDS conditions?  I
: seem to remember some posts about this quite awhile ago, although I think it
: was just IEF and not NEPHGE.  I would apreciate any replies concerning
: incubations in transfer buffer or SDS containing buffer before blotting. 
: Basically, any help would probably be usefull.  

: Chuck Peterson
: Whitney Lab.
: St.Augustine, FL

	Do you mean A) transfering a 1-dimensional NEPHGE gel, or 
                    B) transferring a 2-D, NEPHGE/SDS gel?

	I haven't done A, but it should be like doing an IEF except 
that bands would be in different places.

	"B" is not really any different than a 2D IEF/SDS gels.

	The only thing that might be different is that with prolonged
IEF, you might deplete the ampholytes in the gel, whereas in NEPHGE 
(usually run for a shorter time) you probably haven't lost as much ampho-
lyte during the run.  It just dawns on me now that you might mean 
you are worried about ampholytes binding to nitrocellulose/PVDF/nylon, etc.
In that case, you might have problems.  But let us know what the case is,
and someone "out there" will probably be able to help.....

	good luck, Steve.
--
Steve Driska, Physiology Department, Temple University Medical School
Philadelphia, PA 19140 USA (215) 707-3283 
driska at astro.ocis.temple.edu  
If I could think of something witty and clever to say, it would go right 
here ----->>    .........................