Kendall is absolutely correct.. Faisal
On 3/30/11, Kendall Nettles <knett...@scripps.edu> wrote: > There is a paper from John Kuriyan on co-expressing a phosphatase with c-Src > kinase domain to enable bacterial expression of homogenous protein ( > PMID: > 16260764). Also look at work from E. Goldsmith (PMID: 16829129) > > Lastly, I would suggest general approaches, such as: varying the ends of the > DNA construct; checking different buffers (EDTA, reducing agents, glycerol, > pH, ligands) with use DLS or analytical gel filtration to check for > aggregation. Also try low temp induction, or different fusion proteins. > > Maybe this is a good thing. With our favorite protein we often get > aggregation of half the protein. We assume this is the misfolded protein. We > pellet this and have dozens of structures using the supernatant. So maybe > your aggregation is a feature and not a bug. > > Kendall Nettles > > > On Mar 29, 2011, at 8:10 PM, Neeraj Kapoor wrote: > > Hi All, > I am trying to express a kinase but unfortunately there is aggregation > happening as the protein is purified over a column. SInce I am new to the > field of kinase expression and purification, I was wondering if someone > could provide me with a couple of good references that can hit the ground > running for me. I would also very much appreciate any helpful suggestions > that anyone might have. > > thanks > Neeraj > >
