35S metabolic labeling of ES cells
gilbride at bu.edu
Thu Jan 23 12:35:08 EST 1997
In article <Pine.SUN.3.93.970109170041.4470A-100000 at ruly46>, Peter
Hohenstein <mirihyel at ruly46.medfac.leidenuniv.nl> wrote:
> I'm planning to use 35S metabolic labeled ES (embryonal stemcells -
> murine) cells for an immunprecipitation. Is there anybody having
> experience with labeling of ES cells?
> Thanks for any info
> Peter Hohenstein
> Dept. Human Genetics
> University of Leiden
> The Netherlands
> tel. (+)31 71 5276086
> fax (+)31 71 5276075
I have been metabolically labeling proteins from fetal mouse lung
epithelial cells using using a methionine free medium and 35S labelled
methionine, ~1uCi/ml. Methionine as you know is the initiator amino acid
for nearly all proteins and is probably your best starting point.
Immunoprecipitate (x-body then protein A) from the medium or the cell
lysate, run it, dry it and expose it, your done.
Pretty easy stuff. Good Luck.
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