Has anyone out there had much experience with labelling
Chinese Hamster Ovary cells with 35S-labelled cystein and methionine?
I am wanting to do short labelling times (15 minutes) with CHO cells
but I have been unable to obtain consistent results in my labelling.
What I want to know is do most people pre-incubate CHO cells in a
cysteine and methionine-free medium before adding label? (in order
to reduce intracellular pools of these two amino acids) If so,
how long is a "safe" time without any interuption to protein-synthsis
and hence the viability of the cell?
Also- is such a pre-incubation step necessary for long labelling times?
Thanks in advance,
David Misztal
Dept. Biotechnology
University New South Wales
Australia
