If your protein is His-tagged: spin down cells @~1500xg, adust pH with tris buffer and NaOH to ~pH 7.1 (spin again higher speed if necessary to remove more particulates) and pass over chemical resistant Ni-resin.
Linda Olson, PhD Assistant Professor/ x-Ray Facility Manager Dept. Biochemistry Medical College of Wisconsin 8701 Watertown Plank Rd Milwaukee, WI 53226 phone 414-955-8545 fax 414-456-6510 ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Gloria Borgstahl <gborgst...@gmail.com> Sent: Monday, April 20, 2020 10:56 AM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: [ccp4bb] insect secretion recommendations? ATTENTION: This email originated from a sender outside of MCW. Use caution when clicking on links or opening attachments. ________________________________ Hi Friends, We are secreting Spike Ecto domain into the media from insect cells for purification. As we scale up I am wondering what is recommended for collecting the media from large volumes of culture. Centrifugation? Filtration of some kind? I imagine we need to be gentle to not lyse the insect cells. Thank you, Gloria ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1<https://urldefense.com/v3/__https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1__;!!H8mHWRdzp34!vuXFQA6H16Z3IMEoH8_DuL6_jQcrvgKMWdLAN9ah-kAv8NBJM60Azr55fsNUgg$> ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
