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Protein extraction from gram-positive bacteria

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#1 telma



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Posted 03 July 2002 - 04:26 AM

Any protocols??  I am particularly looking to extract membrane and cytoplasmic fractions that will be resolved on SDS-PAGE. Any suggestions will be most appreciated.

(Edited by telma at 12:09 am on July 3, 2001)

#2 Sciad



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Posted 03 July 2002 - 06:06 AM

You can work with glass beads, size about 1.0 mm.
Mix an equal volume of glass beads with pellet of your bacteria and vortex vigorously.
If available, use an MSK cell hommogeniser from B. Braun Biotech International for this purpose. In this case, you have very strong mixing force combined with the ability to cool your sample during the disruption.
Additional hint:
I recommend to wash the glass beads with your buffer before you first use them. Let me know in case you need more info.
Wishing you success!

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