Dear Cynthia,

I developed a method for lysis by decavitation of nitrogen in a Parr bomb back when I was a post-doc.
This was more efficient and amenable to scale up than the dounce homogenizers that the lab was using.
An additional advantage is that you don't need to use detergent for lysis which can cause problems if internal organelles are also disrupted.  It is well described in Methods in Enzymology 196, 84 - 91 (1991).

Good luck.

regards, marcus


________________________________________
From: DICTY [[log in to unmask]] on behalf of Damer, Cynthia Kay [[log in to unmask]]
Sent: Monday, April 11, 2011 2:06 PM
To: [log in to unmask]
Subject: [DICTY - 542] GST-fusion protein purification from Dicty

Dear Colleagues,

Does someone have a protocol for purifying GST-fusion proteins from
Dictyostelium cells?  I'm most concerned with finding a way to lyse the
cells.  I used to use a French Press, but no longer have access to one.

Thanks,

Cynthia

Cynthia K. Damer
Associate Professor
Biology Department
Brooks Hall Room 229
Central Michigan University
Mount Pleasant, MI 48859
(989) 774-3455

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