Where is Polyacrylamide soluble in?
matthi...@my-deja.com
Can anyone give me some advise, where a polyacrylamide gel is soluble
in? The gel consists of 10 percent polyacrylamide and 90 percent water.
Thanx,
Matthias
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Uncle Al
matthi...@my-deja.com wrote:
>
> Hi!
> Can anyone give me some advise, where a polyacrylamide gel is soluble
> in? The gel consists of 10 percent polyacrylamide and 90 percent water.
Polyacrylamide is soluble in water. That's a short short solvent
list.
If you polymerize acrylamide in methyl methacrylate with N-vinyl
pyrrolidinone to compatiblize the phases you get an intermediate state
when the acrylamide polymerizes and precipitates to create a
magnificent opalescent gel. Then everything polymerizes and you get a
white blob at the end.
--
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matthi...@my-deja.com
>> Can anyone give me some advise, where a polyacrylamide gel is
>> soluble in? The gel consists of 10 percent polyacrylamide and 90
>> percent water.
> Polyacrylamide is soluble in water. That's a short short solvent
> list.
> If you polymerize acrylamide in methyl methacrylate with N-vinyl
> pyrrolidinone to compatiblize the phases you get an intermediate state
> when the acrylamide polymerizes and precipitates to create a
> magnificent opalescent gel. Then everything polymerizes and you get a
> white blob at the end.
and N,N-methylene-bisacrylamide (ratio 29:1), solved in water.
Ammoniumpersulfate was added as initiator and
N,N,N',N'-tetramethylethylenediamin ("TEMED", "TMEDA") as a radical
stabilisator. Polymerisation occured within a few minutes and gave a
white opalescent gel.
My question is where this gel is soluble in.
John Spevacek
> Hi!
> Can anyone give me some advise, where a polyacrylamide gel is soluble
> in? The gel consists of 10 percent polyacrylamide and 90 percent water.
A crosslinked polymer gel is not soluble in anything. Water will swell it,
as will uncrosslinked polymer.
John
--
"A conclusion is the point at which you stopped thinking."
Opinions expressed herein are my own and may not represent those of my employer.
Bam Bam
>
> >> Can anyone give me some advise, where a polyacrylamide gel is
> >> soluble in? The gel consists of 10 percent polyacrylamide and 90
> >> percent water.
> > list.
> > If you polymerize acrylamide in methyl methacrylate with N-vinyl
> > pyrrolidinone to compatiblize the phases you get an intermediate state
> > when the acrylamide polymerizes and precipitates to create a
> > magnificent opalescent gel. Then everything polymerizes and you get a
> > white blob at the end.
> Sorry, perhaps my question was not exact: I had a mixture of acrylamide
> and N,N-methylene-bisacrylamide (ratio 29:1), solved in water.
> Ammoniumpersulfate was added as initiator and
> N,N,N',N'-tetramethylethylenediamin ("TEMED", "TMEDA") as a radical
> stabilisator. Polymerisation occured within a few minutes and gave a
> white opalescent gel.
> My question is where this gel is soluble in.
> Thanx,
> Matthias
A white opalescent gel??? Sounds like you wanted to make a normal gel
for electrophoresis. These gels are notoriously transparent. Unless
there was something really wrong with any of the mixtures. It may
dissolve in a slightly polar solvent (DMSO? DMF? Dioxane?). But why do
you want to dissolve it? Is it stuck somewhere?
Tadeusz Moczon
matthi...@my-deja.com wrote:
> Sorry, perhaps my question was not exact: I had a mixture of acrylamide
> and N,N-methylene-bisacrylamide (ratio 29:1), solved in water.
> Ammoniumpersulfate was added as initiator and
> N,N,N',N'-tetramethylethylenediamin ("TEMED", "TMEDA") as a radical
> stabilisator. Polymerisation occured within a few minutes and gave a
> white opalescent gel.
> My question is where this gel is soluble in.
> Thanx,
> Matthias
========================================
White, opalescent gel? The gel should be perfectly transparent and
non-opalescent!
Well, if you want to recover electrophoresed macromolecules from a
polyacrylamide gel you must either electroelute them from excised gel
fragments or solubilize these fragments. Electroelution calls for the
use of a special apparatus. Solubilization of acrylamide-bisacrylamide
gel is possible but requires the use of a very corrosive agent. If you
wish to obtain easily solubilizable gels, use ethylenediacrylate (=EDIA,
=ethylene glycol dipropenoate) or N,N'diallyltartardiamide (=DATD)
INSTEAD of bis-acrylamide. Look for respective literature.
An acrylamide-ethylenediacrylate polymer serves for electrophoretic
separation at acid, neutral, and only slightly alkaline conditions.
Following separation gel slices can be solubilized by hydrolysis of
ester bonds with alkaline agents:
a) 1 M ammonium hydroxide for a few hours (400 mm^3 of gel per 1 ml of
ammonium hydroxide). Then ammonia can be neutralized or evaporated.
b) Sodium hydroxide soln.
c) A mixture of 1 M piperidine (9 vol) and hyamine_10-X (1 vol) for few
hours at 40o^C.
An acrylamide-N,N'diallyltartardiamide polymer can easily by solubilized
with 2% periodic acid (30-40 min at room temperature).
An acrylamide-bisacrylamide gel is soluble in 6 M HCl, at 120o^C (about
16 hrs, in a Pyrex, fused ampoule). DRIED gel slices one can also
dissolve in 30% hydrogen peroxide (2-6 hrs at 60o^C in a tightly capped
vial).
Good luck.
T.M.
P.S. Elution of macromolecules from frozen, then crushed slices of
AGAROSE gel is very easy. Starch gel slices can easily be solubilized
into maltose and limit dextrins with a trace amount of a pure pancreatic
alpha-amylase at pH 7.6-8.0 or, simply, with few drops of filtered
saliva (at pH 6.5) which contain amylase (you may not use amylase if you
are going to separate proteins). Starch gel offers the lowest
resolution, however.
Frank R. Gorga
In the "old days" when I wanted to count radiolabeled proteins, I would cut
out the band of interest and "dissolve" it by heating at 90 deg C in some
30% H2O2. You can't recover intact protein this way, but it was enough to
get a homogenous sample for liquid scintillation counting.
BioRad used to sell some "fancy" cross linkers that could be used to replace
the bisacrylamide in the standard recipe. These allow one to solubilize the
gel under milder conditions. I don't remember the chemistry involved... it's
been a while.
Hope this helps,
--- FRG
------------------------------------------
Frank R. Gorga, Ph.D.
Dept. of Chemical Sciences
Bridgewater State College
Bridgewater, MA 02325
508-531-2827
508-531-1785 (fax)
fgo...@bridgew.edu
matthi...@my-deja.com wrote in message <8b5035$3pu$1...@nnrp1.deja.com>...
>
>>> Can anyone give me some advise, where a polyacrylamide gel is
>>> soluble in? The gel consists of 10 percent polyacrylamide and 90
>>> percent water.
>> Polyacrylamide is soluble in water. That's a short short solvent
>> list.
>> If you polymerize acrylamide in methyl methacrylate with N-vinyl
>> pyrrolidinone to compatiblize the phases you get an intermediate state
>> when the acrylamide polymerizes and precipitates to create a
>> magnificent opalescent gel. Then everything polymerizes and you get a
>> white blob at the end.
>Sorry, perhaps my question was not exact: I had a mixture of acrylamide
>and N,N-methylene-bisacrylamide (ratio 29:1), solved in water.
>Ammoniumpersulfate was added as initiator and
>N,N,N',N'-tetramethylethylenediamin ("TEMED", "TMEDA") as a radical
>stabilisator. Polymerisation occured within a few minutes and gave a
>white opalescent gel.
>My question is where this gel is soluble in.
>Thanx,
>Matthias
>
>