Interesting 'off-topic' thread. I'm a rather long-time user of Microfluidics cell disruptors (for E. coli or P. pastoris or S. ceresvisiae), and a shared-use M110-P Plug and Play is used by most of the membrane-heads at my place. I've generally been happy (or at least not unhappy) with it.
However, we've been having some QC issues with a membrane protein that we're making in S. ceresvisiae (Sc), and I'm having some concerns about sample heating. Can anyone comment on Microfludics vs Avestin vs Constant Systems vs Retsch vs whatever-else for cracking Sc cells? These days, we're working up ~300-400g of paste at a time. Thank you very much! -MW Michael C. Wiener, Ph.D. Professor Department of Molecular Physiology and Biological Physics University of Virginia PO Box 800886 Charlottesville, VA 22908-0886 434-243-2731 434-982-1616 (FAX) On Wed, 5 Feb 2014 00:34:11 -0500 Anirban Banerjee <ani...@gmail.com> wrote: >I will be curious to know about people's experiences with membrane >proteins and lysing yeast cells with the Microfluidizer and how that >compares with using a Retsch Miller, i.e. grinding in a liquid >nitrogen cooled stainless steel chamber and plunging in liquid >nitrogen in between grinding cycles. > >I am worried that the Microfluidizer is not as mild w.r.t. heating as >they claim it to be. That would, of course, perfectly qualify as my >OCD. > >Any insights will be really appreciated. > >Thanks, > >Anirban > >On Tue, Feb 4, 2014 at 11:58 AM, Matthew Franklin <mfrank...@nysbc.org> wrote: >> Hi Phoebe - >> >> "Cost-effective" may not be the applicable word here, but the Microfluidizer >> works very well: >> >> http://www.microfluidicscorp.com/index.php?option=com_content&view=article&id=19&Itemid=76 >> >> This gadget runs on house compressed air (don't try to use a compressed air >> tank - you'll empty it in minutes). It's a bit noisy, but so is a >> sonicator. >> >> The Microfluidizer really shines with large volumes of lysate - like 1 L and >> up. If you're only processing 100-200 mL at a time, I think sonication is >> the best way to go. >> >> Hope that helps, >> Matt >> >> >> >> On 2/4/14 11:49 AM, Phoebe A. Rice wrote: >> >> Some time ago, there was a nice discussion of cost-effective, wimpy >> protein-friendly ways to break open E. coli. We're thinking about replacing >> an aging sonicator. If people have a favorite gizmo, could they repeat that >> advice? >> thank you, >> Phoebe Rice >> >> ++++++++++++++++++++++++++++++++++++++++++ >> >> Phoebe A. Rice >> Dept. of Biochemistry & Molecular Biology >> The University of Chicago >> >> 773 834 1723; pr...@uchicago.edu >> http://bmb.bsd.uchicago.edu/Faculty_and_Research/ >> >> http://www.rsc.org/shop/books/2008/9780854042722.asp >> >> >> >> -- >> Matthew Franklin, Ph. D. >> Senior Scientist >> New York Structural Biology Center >> 89 Convent Avenue, New York, NY 10027 >> (212) 939-0660 ext. 9374
