Hi Reza, In addition to the many useful suggestions already made, I would suggest lowering the final concentrations of IPTG. In many cases, 1mM IPTG interferes with expression levels and/or solubility. This suggestion does not address your concern for why things become ugly in going from 3mL to 500mL (for which there can be many many explanations), but its worth a try before you head off to make many more clones.
Cheers and best wishes, Raji On Thu, Mar 19, 2015 at 3:33 PM, Reza Khayat <rkha...@ccny.cuny.edu> wrote: > Hi, > > > > We can express quite a bit of soluble protein when growing 3ml cultures. > However, the protein becomes insoluble (inclusion bodies) when we scale up > to 500ml cultures. Has anyone experienced such a problem, and found a > solution to it? Thanks. > > > > Best wishes, > > Reza > > > > Reza Khayat, PhD > > Assistant Professor > > Department of Chemistry > > City College of New York > > New York, NY 10031 > > http://www.khayatlab.org/ > > 212-650-6070 > > >
