[Protein-analysis] Large Scale, native, periplasmic prep

assa sittner via proteins%40net.bio.net (by sittner from lkb.ens.fr)
Fri Sep 12 05:30:00 EST 2008


Hi everyone,
I have been using an osmotic shock (20% sucrose and then water - taken from
"current protocols" or the pET manual) - based protocol, for the extraction
of a protein expressed with pET26b (periplasmic exp. with a pelB leader
sequence).
It looks fine on the small scale (>50ml culture), but I'm having trouble
up-scaling it to ~1L.
does anyone know a protocol designed for large scale periplasmic preparation
(non-danaturing).
cheers,

-- 
Assa Sittner
Optics & Biology group,
Physics Dept.
Ecole Normale Superieure, Paris


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