Hi Matt,
Why not check the NEB pMal system manual
(http://www.neb.com/nebecomm/ManualFiles/manualE8000.pdf). It has a good
cold osmotic shock protocol that I've been using for my periplasmic MBP
fusions.
Zhijie
----- Original Message -----
From: "Matt Colins" <matt.colins2...@yahoo.com>
To: <CCP4BB@JISCMAIL.AC.UK>
Sent: Monday, February 09, 2009 12:52 PM
Subject: [ccp4bb] protocol for harvesting proteins from bacterial
periplasmic space
Dear all,
I plan to prepare a protein fused to pelB signal peptide and secreted to
bacterial periplasmic space. Does anyone have a detailed protocol for
harvesting the protein from bacterial periplasmic space? The protein needs
to be in the native state and will be purified on a nickel column.
I would appreciate your help very much!
Matt
