Help with orthophosphate labeling!

olivier olivier at lan1.molonc.mcgill.ca
Fri Aug 22 09:07:03 EST 1997


Hi,
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
Montreal, Quebec
Canada  H3A 1A1
Phone: (514) 842 12 31 ex: 5832
Olivier at lan1.molonc.mcgill.ca



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