Solubilization of inclusion Bodies

[Ding Liu]

Keywords: 

I need some help about how to complelely solubilze overexpressed protein. I am using PET system to overexpress a DNA binding protein, after cell lysis, protein stays in insoluble fraction, and I tried use 8M urea to solubilize the protein, turns out that more than 250 ml 8M Urea buffer were needed for about 7 g of wet cell, and I still see quite bit protein left in pellet.  Is there a better way to do this, since such large volume is realy not fun to work with ( I am not using His-tag etc. for purification).

Any ideas or suggestions are wellcome.

Thanks in advance.

-- 
Dingjiang Liu
Department of Biochemistry & Biomolecular Structure Analysis Center
University of Connecticut Health Center
263 Farmington Ave., 
Farmington, CT 06032

Phone # 203-679-3117
Fax: 203-679-3408
Email:  DLIU at panda.uchc.edu