Recycling Coomassie destain
Wolfgang Schechinger
Hi.
I suggest distillation of the mix. But I can't warrant that, if you
store the sol'n, that you also will get methyl acetate since MeOH and
HOAc will react (in a chemical equilibrium) to the ester. I only can
say that I used a premixed sol'n stored at RT for more than three
weeks without problems.
You alternatively could switch to a more water soluble dye a ponceau
red, that is easily destained with a buffer containing gelatin or
othe proteins.
Wolfgang
> Dear colleagues,
>
> In order to reduce the amount of chemical waste, our lab would like
> to recycle the MeOH/HAc destain we use for Coomassie stained gels,
> by removing the stain. Has anyone figured out a convenient way of
> doing this? Is activated carbon the best thing to use, or will
> filtration through a thick stack of paper do?
>
> Any suggestions are welcome,
> Thanks,
>
> Marc Werten (m.w.t....@ato.dlo.nl)
> Dept. Microbial Conversions
> Agrotechnological Research Institute (ATO-DLO)
> Bornsesteeg 59
> 6708 PD Wageningen
> The Netherlands
>
> FAX: +31.317.475347
>
>
>
-----
Wolfgang Schechinger
University of Tuebingen
email: wgsc...@med.uni-tuebingen.de
http://www.medizin.uni-tuebingen.de/~wgschech/research.htm
David Micklem
In article <8014181823071997.A07419.ATO4.11B7BC920D00*@MHS>,
M.W.T....@ato.dlo.nl (WERTEN) wrote:
>Dear colleagues,
>
>In order to reduce the amount of chemical waste, our lab would like to recycle
>the MeOH/HAc destain we use for Coomassie stained gels, by removing the stain.
>Has anyone figured out a convenient way of doing this? Is activated carbon the
>best thing to use, or will filtration through a thick stack of paper do?
>
>Any suggestions are welcome,
>Thanks,
>
>Marc Werten (m.w.t....@ato.dlo.nl)
>Dept. Microbial Conversions
>Agrotechnological Research Institute (ATO-DLO)
>Bornsesteeg 59
>6708 PD Wageningen
>The Netherlands
>
>FAX: +31.317.475347
Foam bungs seem to pull out the blue dye pretty well - I used to drop one
in to speed up destaining.
An alternative is to use a colloidal coomassie stain (which is what I do
now). Its available from several people (Integrated Separation Systems
Pro-Blue and Sigma, at least) or you can make it yourself. Then you don't
need to destain, thought the constituents may be worse than MeOH/Acetic
acid: phosphoric acid, ammonium sulphate and MeOH
The original reference is:
Neuhoff V, Arold N, Taube D, Ehrhardt W: Improved staining of proteins in
polyacrylamide gels including isoelectric focusing gels with clear
background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and
R-250. Electrophoresis 1988, 9:255-262.
First heard of it here on methods and reagents....
David
--
D.R.Micklem, Time flies like an arrow...
Wellcome/CRC Institute, Fruit flies like a banana.
Cambridge CB2 1QR, UK Email:dr...@mole.bio.cam.ac.uk
Unsolicited mail will incur a US$100 processing charge.
Holger Steinlechner
WERTEN wrote:
>
> Dear colleagues,
>
> In order to reduce the amount of chemical waste, our lab would like to recycle
> the MeOH/HAc destain we use for Coomassie stained gels, by removing the stain.
> Has anyone figured out a convenient way of doing this? Is activated carbon the
> best thing to use, or will filtration through a thick stack of paper do?
>
> Any suggestions are welcome,
> Thanks,
>
> Marc Werten (m.w.t....@ato.dlo.nl)
> Dept. Microbial Conversions
> Agrotechnological Research Institute (ATO-DLO)
> Bornsesteeg 59
> 6708 PD Wageningen
> The Netherlands
>
> FAX: +31.317.475347
Hi,
we only fill a coloum with some glasfiber in the bottom and put active
carbon on it (about 30-50cm). Then we rinse the destain through the
coloum. That's all and we had no problems till today.
bye
Holger Steinlechner
Hiranya Roychowdhury
At 12:19 AM 7/24/97 -0700, WERTEN wrote:
>Dear colleagues,
>
>In order to reduce the amount of chemical waste, our lab would like to recycle
>the MeOH/HAc destain we use for Coomassie stained gels, by removing the stain.
>Has anyone figured out a convenient way of doing this? Is activated carbon the
>best thing to use, or will filtration through a thick stack of paper do?
>
>Any suggestions are welcome,
>Thanks,
>
>Marc Werten (m.w.t....@ato.dlo.nl)
>Dept. Microbial Conversions
>Agrotechnological Research Institute (ATO-DLO)
>Bornsesteeg 59
>6708 PD Wageningen
>The Netherlands
>
>FAX: +31.317.475347
Cellulosic and polyester materials absorb CBB. For small amounts of the
destain, simply drop a few paper towels or tissues in it and the CBB is
absorbed, leaving behind clean destain. For bulk volumes, you may use
unusable foam(polyester) stoppers or pieces of similar packing materials.
These have almost infinite capacity to absorb CBB. If the storage bottle is
capped securely, such regenerated destains can be used five to ten times.
Near the end, though, you may need do spike it with MeOH.
>
>
>
Dr. Hiranya Sankar Roychowdhury
Plant Genetic Engineering Lab.
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroy...@nmsu.edu
Dr. Duncan Clark
In article <8014181823071997.A07419.ATO4.11B7BC920D00*@MHS>, WERTEN
<M.W.T....@ato.dlo.nl> writes
>In order to reduce the amount of chemical waste, our lab would like to recycle
>the MeOH/HAc destain we use for Coomassie stained gels, by removing the stain.
>Has anyone figured out a convenient way of doing this? Is activated carbon the
>best thing to use, or will filtration through a thick stack of paper do?
We reuse ours several times by adding activated carbon and stirring for
a hour or so (or leave it standing for the odd day or two) then pour the
lot through a column packed with same. We then filter through a couple
of GF/C disks just to remove the last small fines. It gradually goes off
so we make up more etc.
Duncan
--
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk