recovery of proteins from phenol
rfrench at unlinfo.unl.edu
Fri Dec 11 09:54:23 EST 1992
vioque at cica.es (Agustin Vioque) writes:
>I have a highly purified protein that I want to silver stain on a gel. The
>protein is contaminated with a large amount of RNA which will also silver
>stain. Toget rid of the RNA I have tried a treatment with NaOH (10', 80C,
>0.1 N) but a get a smear of partially degraded RNA upon staining of the gel.
>I am thinking of doing a phenol extraction, but how can I recover the protein
>from the phenol phase in a way suitable for SDS-PAGE? If anybody has an
>idea I would appreciate it very much. Sorry for my poor english.
>vioque at cica.es
You can precipitate proteins from phenol by adding 5 volumes methanol
containing 0.1 M ammonium acetate. Hold at -20C for 2 hours,
centrifuge, discard the supernatant, wash with 100% methanol, and dry
in a Speed Vac. The dried pellet is then dissolved in SDS sample buffer.
(procedure courtesy of Les Lane, Univ. Nebraska)
I hope this helps,
Roy French USDA, ARS
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