You can solibilize IB with your solublization buffer, measure the solubilized protein concentration by a protein assay like Bradford. Then adjust the concentration.
Most weight of IBs is not from proteins. However, your protein should be the predominant protein in the IB if the expression is done right. I guess you already have a refolding protocol. Your protein may refold well following the protocol and many host cell proteins don't. You may get rid of many cellular proteins just by doing the refolding. Good luck! Chun Chun Luo, Ph.D. The Protein Expert Accelagen, Inc. 11585 Sorrento Valley Road, Suite 107 San Diego, CA 92121 TeL: 858-350-8085 ext 111 Fax: 858-350-8001 [EMAIL PROTECTED] www.accelagen.com -----Original Message----- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Meg Sent: Monday, July 28, 2008 7:57 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] estimate protein in Incl bod We have our recombinant protein expressed in inclusion bodies we just wash the IBs and perform solubilization and refolding. Our solubilisation protocol mentions making up the volume to contain 7-8 mg / ml protein. 1. how can we determine the total protein content in the inclusion bodies itself which are agregate mass . any good protocol if available. 2. we get our recombinant protein contaminated with host cell potein how to get rid of the host cell proteins. kindly suggest. thanks and regards,
