Re: [ccp4bb] Question about glove boxes for protein crystallization
Just a suggestion. To avoid the glass cover slips, one can use the hanging drop trays from Qiagen (formerly Nextal). No risk of tearing gloves and no grease to mess with either. Crystals can be set up in any glovebox and then the tray will hold the seal using an 0-ring. Nice to be able tomanipulate the crystals in case you have more than one per drop that you want to freeze. Tim Wood > Dear Mathews, > > we were successful in crystallizing two ferredoxins under strict anaerobic > conditions using the much cheaper solution of a glove bag and filling it > with argon. I guess that in both cases (box and bag) the thickness of the > gloves is a problem especially if you are dealing with cover slides. The > glove bag occupies also much less space however its major problem was the > very bad visualization of the inner space and finally we added a plexiglas > window on the bag. Needless to say how essential is not to forget anything > outside the bag/box before starting the crystallization set up. > Finally you can also consider the possibility to use a crystallization > robot. In some of them it is possible to create anaerobic conditions using > argon. > > good luck > Nikos > > > * > Nikos Pinotsis, PhD > EMBL-Hamburg, c/o DESY > Notkestr. 85, Geb. 25A > 22603 Hamburg, Germany > > Phone : +49 40 89902144 > Fax: +49 40 89902149 > e-mail : [EMAIL PROTECTED] > * > > - Original Message - > From: "Mathews, Irimpan" <[EMAIL PROTECTED]> > To: > Sent: Friday, 23 February, 2007 3:03 AM > Subject: [ccp4bb] Question about glove boxes for protein crystallization > > > Dear Friends, > > Sorry for the non CCP4 question. We are planning to purchase a small glove > box to setup crystallization trays under anaerobic conditions. If you have > used glove boxes for crystallization, would you please give me some idea? > > We are thinking of getting the 815 series from Plas-labs (link below). > > http://www.plas-labs.com/ > > Thank you very much, > Mathews > > Ps: If others are interested, I will post a summary. >
Re: [ccp4bb] Question about glove boxes for protein crystallization
Dear Mathews, we were successful in crystallizing two ferredoxins under strict anaerobic conditions using the much cheaper solution of a glove bag and filling it with argon. I guess that in both cases (box and bag) the thickness of the gloves is a problem especially if you are dealing with cover slides. The glove bag occupies also much less space however its major problem was the very bad visualization of the inner space and finally we added a plexiglas window on the bag. Needless to say how essential is not to forget anything outside the bag/box before starting the crystallization set up. Finally you can also consider the possibility to use a crystallization robot. In some of them it is possible to create anaerobic conditions using argon. good luck Nikos * Nikos Pinotsis, PhD EMBL-Hamburg, c/o DESY Notkestr. 85, Geb. 25A 22603 Hamburg, Germany Phone : +49 40 89902144 Fax: +49 40 89902149 e-mail : [EMAIL PROTECTED] * - Original Message - From: "Mathews, Irimpan" <[EMAIL PROTECTED]> To: Sent: Friday, 23 February, 2007 3:03 AM Subject: [ccp4bb] Question about glove boxes for protein crystallization Dear Friends, Sorry for the non CCP4 question. We are planning to purchase a small glove box to setup crystallization trays under anaerobic conditions. If you have used glove boxes for crystallization, would you please give me some idea? We are thinking of getting the 815 series from Plas-labs (link below). http://www.plas-labs.com/ Thank you very much, Mathews Ps: If others are interested, I will post a summary.
Re: [ccp4bb] Question about glove boxes for protein crystallization
Hi Mathews, An anaerobic chamber from Belle Technology (http://www.belle- technology.com/) served me well in the past. Belle's glove boxes are made of acrylic material. That offers relatively low-cost and all-round visibility. Cheers, Roberto On 23 Feb 2007, at 02:03, Mathews, Irimpan wrote: Dear Friends, Sorry for the non CCP4 question. We are planning to purchase a small glove box to setup crystallization trays under anaerobic conditions. If you have used glove boxes for crystallization, would you please give me some idea? We are thinking of getting the 815 series from Plas-labs (link below). http://www.plas-labs.com/ Thank you very much, Mathews Ps: If others are interested, I will post a summary. --- Dr. Roberto Steiner Randall Division of Cell and Molecular Biophysics New Hunt's House King's College London Guy's Campus London, SE1 1UL Phone +44 (0)20-7848-8216 Fax +44 (0)20-7848-6435 e-mail [EMAIL PROTECTED]
[ccp4bb] AW: [ccp4bb] Question about glove boxes for protein crystallization
Dear Patrick, as a (probably more general) follow up-question: Does anyone have experience with batch crystallization under oil for membrane proteins (in detergent) ? alex > Dr. Alexander Pautsch > Protein Crystallography /Structural Research > Boehringer Ingelheim Pharma GmbH & Co. KG Deutschland > Birkendorferstrasse 65 > 88400 BIBERACH, Germany > tel. +49 - (0)7351 - 54 4683 > fax. +49 - (0)7351 - 83 4683 > email [EMAIL PROTECTED] > > -Ursprüngliche Nachricht- Von: CCP4 bulletin board [mailto:[EMAIL PROTECTED] Im Auftrag von Patrick Shaw Stewart Gesendet: Freitag, 23. Februar 2007 10:02 An: CCP4BB@JISCMAIL.AC.UK Betreff: Re: [ccp4bb] Question about glove boxes for protein crystallization Hi Mathews One of our customers, Bret Dillard at the University of Georgia, has had great success with one of our robots in a Bactron X anaerobic chamber. In this case, Bret mainly used the microbatch-under-oil method, which works very well for anaerobic work: (1) the oil protects the sample and reduces exposure to oxygen, (2) the amount of work is far less because you can keep several degassed screens in the chamber to use many times with different protein samples. With vapor diffusion you will have to degass the solutions every for every few samples (if not every sample). Bret won our competition for this work last year. You can find his report at http://www.douglas.co.uk/news.htm, plus more info below Patrick _ Douglas Instruments has announced the winner of the second round of its competition for the best new crystallization technique. Congratulations to Bret Dillard from the University of Georgia, for his winning entry, "Automatic Protein Crystallization in an Anaerobic Environment." Bret placed an Oryx1-6 robot in a Bactron X anaerobic chamber, and used mainly microbatch-under-oil crystallization to crystallize four proteins that are not stable in an environment with oxygen. He found that microbatch avoided the need for frequent degassing of solutions which reduced the work-load enormously. The oil also provided extra protection from oxidation. One of the proteins crystallized is rubrerythrin from the hyperthermophilic archaeon, Pyrococcus furiosus. The native protein contains iron, but is unstable in oxygen. A previously-reported structure was determined in the presence of oxygen, but the iron had been replaced by zinc. Using the anaerobic system, Bret has now obtained the native form containing iron. _ On 2/23/07, Mathews, Irimpan <[EMAIL PROTECTED]> wrote: > > > Dear Friends, > > Sorry for the non CCP4 question. We are planning to purchase a small glove > box to setup crystallization trays under anaerobic conditions. If you have > used glove boxes for crystallization, would you please give me some idea? > > We are thinking of getting the 815 series from Plas-labs (link below). > > http://www.plas-labs.com/ > > Thank you very much, > Mathews > > Ps: If others are interested, I will post a summary. >
Re: [ccp4bb] Question about glove boxes for protein crystallization
Hi Mathews One of our customers, Bret Dillard at the University of Georgia, has had great success with one of our robots in a Bactron X anaerobic chamber. In this case, Bret mainly used the microbatch-under-oil method, which works very well for anaerobic work: (1) the oil protects the sample and reduces exposure to oxygen, (2) the amount of work is far less because you can keep several degassed screens in the chamber to use many times with different protein samples. With vapor diffusion you will have to degass the solutions every for every few samples (if not every sample). Bret won our competition for this work last year. You can find his report at http://www.douglas.co.uk/news.htm, plus more info below Patrick _ Douglas Instruments has announced the winner of the second round of its competition for the best new crystallization technique. Congratulations to Bret Dillard from the University of Georgia, for his winning entry, "Automatic Protein Crystallization in an Anaerobic Environment." Bret placed an Oryx1-6 robot in a Bactron X anaerobic chamber, and used mainly microbatch-under-oil crystallization to crystallize four proteins that are not stable in an environment with oxygen. He found that microbatch avoided the need for frequent degassing of solutions which reduced the work-load enormously. The oil also provided extra protection from oxidation. One of the proteins crystallized is rubrerythrin from the hyperthermophilic archaeon, Pyrococcus furiosus. The native protein contains iron, but is unstable in oxygen. A previously-reported structure was determined in the presence of oxygen, but the iron had been replaced by zinc. Using the anaerobic system, Bret has now obtained the native form containing iron. _ On 2/23/07, Mathews, Irimpan <[EMAIL PROTECTED]> wrote: Dear Friends, Sorry for the non CCP4 question. We are planning to purchase a small glove box to setup crystallization trays under anaerobic conditions. If you have used glove boxes for crystallization, would you please give me some idea? We are thinking of getting the 815 series from Plas-labs (link below). http://www.plas-labs.com/ Thank you very much, Mathews Ps: If others are interested, I will post a summary.
[ccp4bb] Question about glove boxes for protein crystallization
Dear Friends, Sorry for the non CCP4 question. We are planning to purchase a small glove box to setup crystallization trays under anaerobic conditions. If you have used glove boxes for crystallization, would you please give me some idea? We are thinking of getting the 815 series from Plas-labs (link below). http://www.plas-labs.com/ Thank you very much, Mathews Ps: If others are interested, I will post a summary.
