Dear All,
Apologies for the off-topic question. I'm seeking suggestions on the best
way to achieve an effective, but not over-harsh, chemical lysis of E coli
expressing an oxygen-sensitive Fe-S protein. We need to lyse anaerobically
but do not have access to a sonicator that we can use in the glove box. We
have used a proprietary detergent mix in previous attempts but have not
been overly impressed with the results- lots of protein (more than I would
expect based on past experiences with related enzymes) remains in the
pellet. I've seen literature protocols based on Triton (up to 1.2%) but am
worried (perhaps without basis) that this might interfere with downstream
steps (reconstituting the Fe-S cluster and possibly crystallisation). The
protein is His-tagged.
Does anyone know a better way?
Thanks in advance.
Best wishes
Jim
----------------------
Dr. James Spencer,
Lecturer in Microbiology
School of Cellular and Molecular Medicine
Medical Sciences Building
University of Bristol
University Walk
Bristol BS8 1TD
jim.spen...@bristol.ac.uk
http://www.bristol.ac.uk/cellmolmed/staff/spencer.html
----------------------
Tel: (44) (0) 117 331 2084
Fax: (44) (0) 117 331 2091
