Inclusion bodies centrifugation

megha goyal via methods%40net.bio.net (by mgbiotec from gmail.com)
Tue Oct 27 23:16:59 EST 2009


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.

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


On 10/27/09, megha goyal <mgbiotec from gmail.com> wrote:
>
> 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.
>


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