Solubilizing bisacrylamide-crosslinked polyacrylamide gels

Dima Klenchin klenchin at macc.wisc.edu
Thu Jun 20 21:54:04 EST 1996


In article <DtB2oF.1wF at uns.bris.ac.uk>, Mike Perry <Mike.Perry at bris.ac.uk> wrote:
#I'm chopping up a polyacrylamide (10:2.5) gel containing 35S labelled 
#samples to count in a scint counter.  Now Hames and Rickwood (practical 
#approach guide) reckon it's best to solubilize in 30% H2O2 at 50 deg C. 
#then either add Amersham's solubilizer + toluene scint fluid or use a 1:2 
#solution of Triton X-100 and toluene.
#What I'm wondering is whether I can get away with simply solubilizing in 
#H2O2 and then adding my standard aqueous scint (Packard "Emulsifier Safe" 
#and LSC cocktail).  Anyone got any ideas?

Don't know about S35 - I did P32. Hardly there is any difference. As far 
as dissolving PAAG goes, < 2 hr 60C or even rt overnight in 30% 
H2O2 dissolve it completely. Solubilizer is totally unnecessary. All modern
LSCs contain nonionic detergents, so Triton is not needed either. The
only thing that is going to affect you is whether given LSC is mixable with
~ 1/5 - 1/10 part 30% H2O2. This is easy to test. I used Aquasol II from NEN
with excellent results (it's not rated "biodegradable" but can take up to 
1:1 volume of aqueous solutions).

It's important to make sure that H2O2 is fresh. 

Good luck,

- Dima




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