making a master mix for SDS-PAGE

Michelle Gleeson Michelle.Gleeson at uts.edu.au
Tue Nov 7 16:58:41 EST 2000


Many years back, when I did manual sequencing :-( , we routinely made 
up batches of the buffer/water/acrylamide mix. It was kept for no 
longer than one month, at 4degC, in a dark bottle wrapped in foil.  We 
let it warm up a bit, added the TEMED and APS, and poured.  If it is 
too cold when poured, it takes too long to set.  Conversely, when you 
have to make a fresh batch in a hurry, pouring even slightly warm 
resulted in catastrophic  premature setting.  I don't know for sure 
what would happen if SDS were added to the mix, but I imagine it might 
precipitate out at cold temps.  Thus tempting you to warm it up before 
pouring... ;-)

Why not test it out?  Not really much to lose.
Cheers,
Michelle 

Molecular Parasitology Unit
University of Technology, Sydney
'don't cut too many corners, or you'll end up going around in circles'

----- Original Message -----
From: mlsulliv at facstaff.wisc.edu ("Michael L. Sullivan")
Date: Wednesday, November 8, 2000 4:50 am
Subject: Re: making a master mix for SDS-PAGE

> Maybe not.  I just found the following in CSH press "Antibodies: A
> laboratory Manual":
> 
> "For greter reproducibility from day to day, the acrylamide,
> bis-acrylamide, buffer and water can be prepared in lagre batches and
> either stored at 4 deg. C for 1 month or frozen in aliquots and used
> indefinitely.  Remove the required amount, warm to room 
> temperature and add
> ammonium persulfate and TEMED immediately before use."
> 
> I'd still be interested to know if anybody is doing this 
> routinely.  I sure
> hate mixing up gel mix all the time!
> 
> Mike
> 
> >I think it is not done this way because it does not work... 
> otherwise,, we
> >would be doing it this way.
> >
> >"Michael L. Sullivan" wrote:
> >
> >> Since I mostly run 10% SDS-PAGE gels, is there any reason I 
> can't make up a
> >> master mix of acrylamide, water, SDS, and buffer that would 
> need only to
> >> have APS and TEMED added?
> >>
> >> I know that such mixes are available commercially, but I wasn't 
> sure>> whether those might have some additive that would prevent 
> some dire
> >> consequence of mixing the other components for longer term 
> storage and use.
> >>
> >> Thanks for any insight.
> >>
> >> Mike
> >>
> >> Michael L. Sullivan, Ph.D
> >>
> >> U.S. Dairy Forage Research Center
> >> 1925 Linden Drive West
> >> Madison WI, 53706
> >>
> >> (608) 264-5144 Phone
> >> (608) 264-5147 Fax
> >>
> >> ---
> 
> 
> ---
> 
> 
> 


---






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