We have been using Strep-Tactin SuperFlow (high capacity) columns more than 100 times, even when run under preparative conditions with protein extracts from E. coli fermentations.
Biotin binding to the engineered streptavidin is not irreversible but can be cured by extended washing. To check the column status I recommend our HABA binding procedure described in: Schmidt, T. G. M. & Skerra, A. (2007) The Strep-tag system for one-step purification and high affinity detection or capturing of proteins. Nat. Protoc. 2, 1528-1535. Good luck, Arne Skerra Am 11.07.2011 um 22:56 schrieb D Bonsor: > Does anyone know how many times roughly you can re-use a Streptactin column? > I know that contamination with biotin will destroy the column but the protein > that I am using has gone through both a GST and Nickel purification steps > before seeing the Streptactin column and I think lately that I have been > seeing decreased binding. > > Thanks in advanced. + Prof. Dr. Arne Skerra + Lehrstuhl f. Biologische Chemie + Technische Universitaet Muenchen Emil-Erlenmeyer-Forum 5 + 85350 Freising-Weihenstephan + Germany Phone: +49 (0)8161 71-4351 + Fax: -4352 http://www.wzw.tum.de/bc + eMail: ske...@tum.de
