I am working on a thermophilic protein overexpressed in E.coli. The first step in purification is heating. I used Biorad and BCA assay to determine the protein concentration, but the result from the two assay is very different. Protein concentration determined by Biorad is 2X higher. I also have trouble purify the protein, and concentrate the protein to more than 1.5mg/ml. Does this all point to misfolded protein?
http://biowww.net/mynews/tree.php?group_name=bionet_molbio_proteins&begin=0
