inclusion bodies, purification of

slavemaster un691cs at genius.embnet.dkfz-heidelberg.de
Tue Dec 3 05:55:54 EST 1996


In article <32A2F2D4.4E89 at snoopy.org.chemie.tu-muenchen.de>, 
schott at snoopy.org.chemie.tu-muenchen.de says...


>Does anybody know a simple method, (or a literature citation) for 
>separating inclusion bodies from bacterial debris? (probably by 
>ultracentrifugation?)
>
>Thanks 
>
>Karin Schott

Try to get a protocol booklet from either Novagen or Qiagen: 
they describe methods how to purify His-Tagged proteins from
inclusionbodies.

In principal, I spin the E. colis at 5 K, then dissolve the
pellet in PBS, Then give them 5 * 10 seconds on ice-water (!) 
at fullblast setting of ultrasound. Then spin again at 5k.
The pellet now contains bacterial debris. 
Spinning at 13k results in a white pellet of inclusion
bodies that then can be opened with high Urea (6M). 

clemens 
ANAT3,
Heidelberg




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