Thanks a lot for all your suggestions. But how much DTT or BME shall i add. Also i fear that these will interfere in the formulation and how shall i get rid of them then. As for GPC my protein is 19KDa and diemr will be approx 38 KDa and it is difficult to separate them without losing the protein so i am searching for alternative method.
thanks. meg On 2/18/09, Anthony Addlagatta <anth...@iict.res.in> wrote: > > Why don't you add some reducing agent like DTT or BME to make everything as > monomer? > > Anthony > > On Tue, 17 Feb 2009 10:47:28 +0000, Meg wrote > > I have purified my protein granulocyte colony stimulating factor using > > chromatography steps. my protein is relatively pure when analysed by > > reducing and non-reducing SDS PAGE method. however non-reducing page > > shows DIMER presence. i have about 250 ml pure protein sample and do not > > want to perform gel permeation chromatography as it will be time > consuming. > > is there a way to get rid of dimers by some other method. can dialysis > help > > me. kindly guide me as i have to formulate the product and fill it > quickly. > > > > Thanks in anticipation. > > > > meg > > > ------------------------------------------------- > Anthony Addlagatta, Ph.D. > Ramanujan Fellow and Senior Scientist > Center for Chemical Biology > Indian Institute of Chemical Technology [IICT] > Tarnaka, Hyderabad- 500007, INDIA > Tel:+91-40-27191583 > Url: http://www.iictindia.org/zacb/Dr.%20Anthony.aspx > >
