Protein precip. from Guanidine
nicholas_theodorakis at urmc.rochester.edu
Tue Oct 12 22:47:41 EST 1999
In article <3803d94b.356044020 at news>,
jrt at home.com (John Thompson) wrote:
> Kresten <kresten at my-deja.com> wrote:
> >Could anybody please suggest a method for precipitation of protein
> >subsequent SDS-PAGE.
> >My sample volume is 200 microliters and contains 40 micrograms of
> >protein). Furthermore the sample is 6.8M in Guanidine Hydrochloride.
> Why precipitate at all. You can probably add 5X sample buffer and load
> the sample directly on SDS Page. If you need to concentrate to I'd
> suggest a low mw cutoff spin filter (try Amicon or Millipore) that
> will allow you to change the buffer and reduce the volume at the same
> John Thompson
> Merck Research Labs
The problem will be the ionic content of the guanidine, which will
surely mess up the electrophoresis. It wouldn't be a problem with urea
(but I would skip the boiling step). Is there a way the the original
poster could dialyze against urea to remove the guanidine (assuming a
denaturant was needed to keep it in solution)?
| Nick Theodorakis |
| nicholas_theodorakis at urmc.rochester.edu |
| (previously theodorn at gusun.georgetown.edu) |
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