Radiolabelling proteins in Coli

john brennand john.brennand at gbapr.zeneca.com
Mon Apr 15 09:49:23 EST 1996


We are having trouble visualising a recombinant protein on coomassie 
gels. So, has anyone got a good "money back" protocol for radiolabelling 
newly synthesised proteins in E Coli following induction (iptg) of 
expression from a plasmid ?

What I need to know are things like:

Growth state of the cells, media composition, which radiolabel works 
best, how much label to use, time of incubation, etc.

I guess I could do experiments to work all of this out but I just want a 
quick answer to whether our construct is making any recombinant protein 
and if so, is it being degraded intracellularly

Thanks for any help

John





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