Bacterial expression: incorporating radioactivity into overexpressed protein - (Nov/04/2011 )
I'd like to incorporate radioactive amino acids into a protein that's overexpressed in bacteria upon addition of IPTG. It doesn't really matter to me whether the hot amino acids are 35-S or 14-C, the most important thing is that the protein is as hot as possible so that it can be detected readily by autoradiography. So the questions:
1) Can anybody point me in the direction of some type of dropout medium that can be supplemented with hot amino acids?
2) Has anybody done such an experiment and found a high degree of radiolabel incorporation?
3) Does anybody have a reference to a paper that achieved the above?
Many thanks in advance.
there is a system available from takara, especially for protein labeling! ...i would go for such a system because otherwise your precious labled aminoacids are wasted on all the other E. coli proteins!
You can find the details here!