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Large Scale, native, periplasmic prep

Large Scale, native, periplasmic prep - Protein Forum

Large Scale, native, periplasmic prep - Protein Forum


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Old 09-12-2008, 10:30 AM
assa sittner
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Default Large Scale, native, periplasmic prep



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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