Help with orthophosphate labeling!
olivier at lan1.molonc.mcgill.ca
Fri Aug 22 09:07:03 EST 1997
I'm trying to label a protein in vivo with orthophosphate. Results are
not great so far. I've done a 7 hours incubation with 8 mCi (0.8
mCi/ml), with something like 10 millions NIH 3T3 cells (around 1 mg of
protein at the end), followed by IP and 5 washes with the extraction
buffer. Still, I got a lot of background and a very weak band.
Is it worth trying a longer incubation with 32P, or starting with more
cells? How can I remove the background?
Any idea would be very helpful!
Olivier Coqueret. Ph. D.
Molecular Oncology Group
McGill University, Royal Victoria Hospital
687 Pine Av. West
Canada H3A 1A1
Phone: (514) 842 12 31 ex: 5832
Olivier at lan1.molonc.mcgill.ca
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