- try limited proteolysis to see if you can chop off a disordered region - consider the fact that, although it purifies nicely, your protein may not be well-folded do you have a biochemical activity test?
Mark J van Raaij Laboratorio M-4 Dpto de Estructura de Macromoleculas Centro Nacional de Biotecnologia - CSIC c/Darwin 3, Campus Cantoblanco E-28049 Madrid, Spain tel. (+34) 91 585 4616 http://www.cnb.csic.es/content/research/macromolecular/mvraaij/index.php?l=1 On 23 Mar 2011, at 18:59, Van Den Berg, Bert wrote: > Try different detergents. > Try 10% or more glycerol. > Try adding ligands (if present/known). > Try varying ionic strength and/or pH. > Try giving more specifics so people on the board may be able to help you > better. > > Bert > > > On 3/23/11 1:51 PM, "gauri misra" <kamga...@gmail.com> wrote: > > The protein purifies nicely there is no problem in that. Just at the last > step when it is concentrated it starts precipitating beyond a concentration > of 1mg/ml. > Already the purification buffers have the detergent. > > On Wed, Mar 23, 2011 at 1:44 PM, Kornelius Zeth > <kornelius.z...@tuebingen.mpg.de> wrote: > > 2 M urea and detergents > > On Wed, 23 Mar 2011 13:41:55 -0400 > gauri misra <kamga...@gmail.com> wrote: > > Hi, > > What are the different methods to prevent protein aggregation while > > concentrating so as to increase the concentration of the protein? > > I have some idea of adding EDTA and charged amino acids like L-Arg and > > L-Glu. > > I would appreciate if the readers share their experiences. > > > > Thanks! > > > > Gauri > > ---------------------------------------------- > Kornelius Zeth > Max Planck Institute for Developmental Biology > Dept. Protein Evolution > Spemannstr. 35 > 72076 Tuebingen, Germany > kornelius.z...@tuebingen.mpg.de > Tel -49 7071 601 323 > Fax -49 7071 601 349 > >
