NEPHGE and SDS transfer

Stephen P. Driska PhD driska at
Wed Jan 3 14:34:25 EST 1996

rep at 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  
If I could think of something witty and clever to say, it would go right 
here ----->>    .........................

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