At 04:51 PM 8/31/1999 +0200, fred wrote:
>I run SDSPAGE to separate two proteins.Then I want to elute one of
>them.Does anybody know simple way to extract protein from acrylamide gel?
>thank you,
Fred,
We find passive extraction to be simple and effective. "Effective" hasn't
been exhaustively documented, but it was greater than 50% for a selenium-75
labelled protein with this method. We got the "recipe" from
Hewlett-Packard, and I think it's published in one of their application
notes (AN 934 ?):
1. Coomassie staining does not interfere with sequencing, so go ahead
and stain.
2. Cut out the desired band(s) and place in a Sarstedt or Eppendorf
tube. Mince the gel with forceps.
3. Wash the gel 3 times with 1.0 ml water, discarding the supernatant
each time.
4. Add a maximum of 0.50 ml (preferably less) 100 mM TrisHCl, pH 8.5,
with 0.1% SDS, vortex well, and allow to stand overnight.
5. Centrifuge and save the supernatant.
6. Add another 0.25 ml (maximum) of the Tris/SDS, vortex, and
centrifuge. Combine the supernatant with the first 0.5 ml.
That's all it takes.
Rod Levine
NIH
Bldg 3, Room 106 MSC 0320
Bethesda, MD 20892-0320
email: rlevine@nih.gov
voice: 1 (301) 496-2310
fax: 1 (301) 496-0599
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At 04:51 PM 8/31/1999 +0200, fred wrote:
I run SDSPAGE to separate two proteins.Then I
want to elute one of
them.Does anybody know simple way to extract protein from acrylamide
gel?
thank you,
Fred,
We find passive extraction to be simple and effective.
"Effective" hasn't been exhaustively documented, but it was
greater than 50% for a selenium-75 labelled protein with this
method. We got the "recipe" from Hewlett-Packard, and I
think it's published in one of their application notes (AN 934 ?):
1.
2.
3.
4.
5.
6.
That's all it takes.
Rod Levine
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