Quick SDS PAGE question

Andrew Hobbs andrewh at uniwa.uwa.edu.au
Thu Jul 29 21:38:34 EST 1993


Sarah Tilley (sjt at newt.phys.unsw.edu.au) wrote:

: --
: Hi,

: I have a recipe for a 10% SDS gel (supplied by a company) which seems
to take to long to polymerise according to my previous experiences with
SDS PAGE. When I looked at the recipe, the amount of TEMED seems quite
low- 20ul of TEMED and 300ul of 10% AMPS per 60 ml of separating gel.
Would I mess up the gel if I increased the TEMED to 50 or 60ul? I have
a separate recipe for a 15% gel that uses 170ul TEMED and 450ul of 10%
AMPS for 50 mls. Will playing around with these chemicals affect
anything else other t

: han rate of polymerisation?

: Thanks in advance
: **
: Sarah Tilley
: sjt at newt.phys.unsw.edu.au

In my experience 20 ul TEMED is plenty for a 60 ml gel.  I tend to use 5
- 10 ul.  The degradation products of TEMED (ie from old TEMED) can in
fact inhibit the reaction.  If the gel is taking too long to set 
generally I immediately suspect the degassing was not sufficient or
alternatively the ammonium persulphate has gone off (We did have one
bottle of Ammonium persulphate which resolutely refused to cause
polymerization of acrylamide though the bottle was several years old so
I didn't complain)

Andrew Hobbs
Biochemistry
University of Western Australia

andrewh at uniwa.uwa.edu.au



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