Dear colleagues, The CCP4BB community is amazing, as always. I received many comments by happy users of the Avestin Emulsiflex C3 (and C5); not so much on the Microfluidics LM20. Thanks for that, it will be a great help to decide which instrument to purchase.
Best, Bernhard From: CCP4 bulletin board <[email protected]> on behalf of Adam Middleton <[email protected]> Reply to: Adam Middleton <[email protected]> Date: Tuesday, 18 August 2020 at 2:56 pm To: "[email protected]" <[email protected]> Subject: Re: [ccp4bb] Cell disruption Hi Bernhard, The Department has an Avestin emusiflex here, and it is kept in the cold room. Works great as long as your cells are properly resuspended and in a big enough volume. Highly recommended. Adam On Sun, Aug 16, 2020 at 4:48 PM Pascal Egea <[email protected]<mailto:[email protected]>> wrote: Dear Bernhard , I would recommend the emulsified from Avestin. It is great . Depending on your budget you can ask for stainless steel (probe to slow tear ) or ceramic (longer lasting but a bit more expensive). We have the ceramic but I have worked with the stainless steel one and it is really good with E. coli cells. Not the best for Yeast despite pressurizing more. Yeast are better dealt with bead beaters or cryo-mill grinders. 3 passes are usually sufficient to process up to 250 ml of extract in a decent time (15-20) minutes . We operate ours at room temperature just cool the serpentine tubing coming out of the chamber in ice . There is an option to add a cooling plate but we have never considered it. Overheating comes from overpressurizing but for E Coli at 15000 psi max 3 passes will do the trick without overhea tu bf the sample. I have processEs successfully many membrane proteins And protein complexes with it. I have never seen one in a cold room to be honest but it should be fine although I don’t think it would be necessary . I would Ask avestin about that . Maintenance and cleaning is simple . Once in a while I will replace the seal, it s not very difficult to do. The whole assembly is metal so you can sterilize the whole thing After complete disassembly in extreme cases . This is a bit more involved but not overwhelming. The people at Avestin are super nice and responsive ( Canadians) so they will guide you if necessary . And we have had their engineers show up sometimes to just check the instrument for free . I have had this one for 10 years in my lab now. And before that was using one for 7 years during my post doc. I would not lyse bacteria by any other method now. I hope this helps. All the best. Pascal On Sat, Aug 15, 2020 at 9:08 PM Bernhard Lechtenberg <[email protected]<mailto:[email protected]>> wrote: Dear colleagues, We are currently looking to purchase a cell disruptor/homogeniser mainly for routinely processing a few 100 mls of E. coli suspensions. With the current COVID-19 restrictions it is very difficult for us to test any equipment. I thus hope that some of you can share their experiences with the different models. I found a similar thread on the CCP4BB from 2013 but wondered if anybody had had some more up to date information. We are mainly looking at the Avestin Emulsiflex C3 homogeniser and the Microfluidics LM20 Microfluidizer. In particular we are interested to know more about ease of use, maintenance, reliability and if anybody operates these in a cold room (4°C). Thanks in advance, Bernhard -- Bernhard C. Lechtenberg, Ph.D. Laboratory Head Ubiquitin Signalling Division The Walter and Eliza Hall Institute 1G Royal Parade<https://www.google.com/maps/search/1G+Royal+Parade+%0D%0A+Parkville+VIC+3052+%0D%0A+Australia?entry=gmail&source=g> Parkville VIC 3052<https://www.google.com/maps/search/1G+Royal+Parade+%0D%0A+Parkville+VIC+3052+%0D%0A+Australia?entry=gmail&source=g> Australia<https://www.google.com/maps/search/1G+Royal+Parade+%0D%0A+Parkville+VIC+3052+%0D%0A+Australia?entry=gmail&source=g> Phone: +61 3 9345 2217 Email: [email protected]<mailto:[email protected]> _______________________________________________ The information in this email is confidential and intended solely for the addressee. You must not disclose, forward, print or use it without the permission of the sender. The Walter and Eliza Hall Institute acknowledges the Wurundjeri people of the Kulin Nation as the traditional owners of the land where our campuses are located and the continuing connection to country and community. _______________________________________________ ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 -- Pascal F. Egea, PhD Associate Project Scientist UCLA, David Geffen School of Medicine Department of Biological Chemistry Boyer Hall room 356 611 Charles E Young Drive East Los Angeles CA 90095 office (310)-983-3515 lab (310)-983-3516 email pegea at mednet.ucla.edu<http://mednet.ucla.edu> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 -- Adam Middleton, Ph.D. Research Fellow Department of Biochemistry School of Biomedical Sciences University of Otago 710 Cumberland St. Dunedin 9016 New Zealand [email protected]<mailto:[email protected]> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 _______________________________________________ The information in this email is confidential and intended solely for the addressee. You must not disclose, forward, print or use it without the permission of the sender. The Walter and Eliza Hall Institute acknowledges the Wurundjeri people of the Kulin Nation as the traditional owners of the land where our campuses are located and the continuing connection to country and community. _______________________________________________ ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
