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Inclusion bodies centrifugation
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megha goyal
Oct 27, 2009, 11:18:33 PM10/27/09
to met...@magpie.bio.indiana.edu
Dear All,
Our protein is expressed as inclusion bodies and I want to separate
inclusion bodies from E.coli from the cellular debris after* *lysis of the
cells by sonication.
Can I do this by normal centrifugation? and if yes, at what speed?
Our centrifuge has maximum speed of 14000 rpm. Can we do the separation
using this centrifuge and if so how.
Thanking in anticipation.
megha goyal
Oct 28, 2009, 12:16:59 AM10/28/09
to met...@magpie.bio.indiana.edu
Thanks for the reply. actually we do not use any lysis buffer just suspend
our e.coli cells in water and perform sonication thereafter we centrifuge at
140000 rpm to collect the inclusion bodies and these are contaminated with
host cell proteins too.
our e.coli cells in water and perform sonication thereafter we centrifuge at
140000 rpm to collect the inclusion bodies and these are contaminated with
host cell proteins too.
as mentioned by artem, i just wanted to know if we centrifuge at 5000 - 6000
rpm then will the cell debris not settle down. kindly guide.
as such i shall try the method and revert back. thanks for the reply
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