Solubilization of inclusion Bodies
dliu at panda.uchc.edu
Mon Mar 20 12:20:49 EST 1995
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.
Department of Biochemistry & Biomolecular Structure Analysis Center
University of Connecticut Health Center
263 Farmington Ave.,
Farmington, CT 06032
Phone # 203-679-3117
Email: DLIU at panda.uchc.edu
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