Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Thank you for your comments everyone. The CCP4BB is a wonderful resource and it has answered several questions that have been bothering me for years! Tristran has given us the correct conclusion as well as the important facts: the capacity of oil for holding O2 is high, but the diffusion rate is low. This makes complete sense of the observations reported. The O2 is (slowly) diffusing OUT of the permanganate drop, and the oil is already saturated with O2, therefore it takes a long time for the purple colour to be lost. The O2 is diffusing INTO the drop in the dithionite experiment, and presumably the oil that Julia used was already loaded with O2, so the reducing environment was quickly lost. I hadn't figured out how to take advantage of the protection of oil - I just had a vague feeling that it might be helpful. Now however I can see that it's useful, because the oil will provide quite good protection against a pulse of O2 e.g. if someone accidentally lets air into the chamber. (Or moves plates from one glovebox to another?) O2 will start to diffuse into the oil, but most of it will diffuse out again if the O2 pulse is short. And the lids that are standard on microbatch plates will help a lot. (The oil is almost the ideal barrier, although you *might* prefer something with a very low solubility since it *might *give a lower O2 flux in the steady state - as Tristran says it's complicated. And I'm guessing that the O2 flux through the thin plastic tape used in vapor diffusion setups would be quite high. Does anyone have a friend who works in food science?) There's an even more exciting conclusion: we should degass our oil even for use with *aerobic *microbatch setups. I have heard of a case where diffracting crystals were only obtained for aerobic targets in a glovebox, and I think the skins on drops are, or can be, related to oxidation. There may even be mileage in microbatch with the zip lock bag approach for targets that are not overly sensitive - *if* you degas your oil before you start a vacuum should do it. I agree that Al's Oil (silicone) should be avoided from this point of view - although I would certainly use it anyway for screening experiments (whether aerobic or anaerobic). Riveting stuff. Thx to all, Patrick On 18 March 2015 at 18:58, Tristan Croll tristan.cr...@qut.edu.au wrote: It's a little complicated. It's true that oxygen is more soluble in most oils than in water - but in a high viscosity mineral oil the diffusion rate is orders of magnitude lower. So the combination of an oil overlay and a reducing agent in your buffer should protect your sample much longer than the reducing agent alone - as long as your oil was degassed to start with. Note that silicon oils are a bad choice for this - silicones have an enormous affinity for oxygen (so much so that they've been explored as artificial blood substitutes), and it diffuses through them very readily. Tristan Croll Lecturer Faculty of Health School of Biomedical Sciences Institute of Health and Biomedical Engineering Queensland University of Technology 60 Musk Ave Kelvin Grove QLD 4059 Australia +61 7 3138 6443 This email and its attachments (if any) contain confidential information intended for use by the addressee and may be privileged. We do not waive any confidentiality, privilege or copyright associated with the email or the attachments. If you are not the intended addressee, you must not use, transmit, disclose or copy the email or any attachments. If you receive this email by mistake, please notify the sender immediately and delete the original email. On 18 Mar 2015, at 11:49 pm, Edward A. Berry ber...@upstate.edu wrote: Do you have evidence that the oil blocks diffusion of O2? O2 is a nonpolar molecule, generally much more soluble in oils than in water. I'm not sure about silicone oils, but I would think they also dissolve O2 readily. eab On 03/18/2015 08:02 AM, Patrick Shaw Stewart wrote: Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Do you have evidence that the oil blocks diffusion of O2? O2 is a nonpolar molecule, generally much more soluble in oils than in water. I'm not sure about silicone oils, but I would think they also dissolve O2 readily. eab On 03/18/2015 08:02 AM, Patrick Shaw Stewart wrote: Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk tel 01235 567717 tel:01235%20567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm. -- patr...@douglas.co.uk mailto:patr...@douglas.co.ukDouglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Peter Baldock, Patrick Shaw Stewart http://www.douglas.co.uk Tel: 44 (0) 148-864-9090US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm . -- patr...@douglas.co.ukDouglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Peter Baldock, Patrick Shaw Stewart http://www.douglas.co.uk Tel: 44 (0) 148-864-9090US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Hi, As far as I can tell oil does not block diffusion of O2 whatsoever. You can keep larger volumes (≥1 ml) of solutions anoxic in air for several hours with dithionite (≥0.5%) to scavenge oxygen and a redox indicator dye such as phenosafranin to monitor the state of the solution. Small drops (large surface/volume ratio) however oxidize within seconds, whether or not they are covered with oil. Of course this may simply be because the oxygen gets in before the drop is covered with oil, but either way I don't see how you could set up anaerobic drops in an aerobic environment. Best, Julia On 18/03/15 14:47, Edward A. Berry wrote: Do you have evidence that the oil blocks diffusion of O2? O2 is a nonpolar molecule, generally much more soluble in oils than in water. I'm not sure about silicone oils, but I would think they also dissolve O2 readily. eab On 03/18/2015 08:02 AM, Patrick Shaw Stewart wrote: Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk tel 01235 567717 tel:01235%20567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm. -- patr...@douglas.co.uk mailto:patr...@douglas.co.uk Douglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Peter Baldock, Patrick Shaw Stewart http://www.douglas.co.uk Tel: 44 (0) 148-864-9090US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36 -- Dr. Julia Griese Postdoctoral Researcher Stockholm Center for Biomembrane Research Department of Biochemistry and Biophysics Stockholm University 106 91 Stockholm Sweden phone: +46-(0)8-162 778 email: gri...@dbb.su.se
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
I also wondered about the statement about oils blocking diffusion of O2. We had lots of trouble keeping things anaerobic in a glove box until we degassed the oils and waxes used to mount crystals in capillaries. We found that putting them under vacuum removed much of the dissolved oxygen. The waxes required cycling between heating and vacuum several times. Ron On Wed, 18 Mar 2015, Edward A. Berry wrote: Do you have evidence that the oil blocks diffusion of O2? O2 is a nonpolar molecule, generally much more soluble in oils than in water. I'm not sure about silicone oils, but I would think they also dissolve O2 readily. eab On 03/18/2015 08:02 AM, Patrick Shaw Stewart wrote: Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk tel 01235 567717 tel:01235%20567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm. -- patr...@douglas.co.uk mailto:patr...@douglas.co.ukDouglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Peter Baldock, Patrick Shaw Stewart http://www.douglas.co.uk Tel: 44 (0) 148-864-9090US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
It's a little complicated. It's true that oxygen is more soluble in most oils than in water - but in a high viscosity mineral oil the diffusion rate is orders of magnitude lower. So the combination of an oil overlay and a reducing agent in your buffer should protect your sample much longer than the reducing agent alone - as long as your oil was degassed to start with. Note that silicon oils are a bad choice for this - silicones have an enormous affinity for oxygen (so much so that they've been explored as artificial blood substitutes), and it diffuses through them very readily. Tristan Croll Lecturer Faculty of Health School of Biomedical Sciences Institute of Health and Biomedical Engineering Queensland University of Technology 60 Musk Ave Kelvin Grove QLD 4059 Australia +61 7 3138 6443 This email and its attachments (if any) contain confidential information intended for use by the addressee and may be privileged. We do not waive any confidentiality, privilege or copyright associated with the email or the attachments. If you are not the intended addressee, you must not use, transmit, disclose or copy the email or any attachments. If you receive this email by mistake, please notify the sender immediately and delete the original email. On 18 Mar 2015, at 11:49 pm, Edward A. Berry ber...@upstate.edu wrote: Do you have evidence that the oil blocks diffusion of O2? O2 is a nonpolar molecule, generally much more soluble in oils than in water. I'm not sure about silicone oils, but I would think they also dissolve O2 readily. eab On 03/18/2015 08:02 AM, Patrick Shaw Stewart wrote: Hi Steve I have one more comment for this thread. The microbatch-under-oil method is very handy for anaerobic work: 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. Best wishes Patrick On 11 March 2015 at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk mailto:stephen.c...@rc-harwell.ac.uk tel 01235 567717 tel:01235%20567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm. -- patr...@douglas.co.uk mailto:patr...@douglas.co.ukDouglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Peter Baldock, Patrick Shaw Stewart http://www.douglas.co.uk Tel: 44 (0) 148-864-9090US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Actually I may have misunderstood the original post. Patrick never said oils block O2 diffusion: On 03/18/2015 09:47 AM, Edward A. Berry wrote: The microbatch-under-oil method is very handy for anaerobic work: (In a glove box, of course) 1. You can keep the microbatch stock solutions in normal microtitre plates (polypropylene is best to reduce evaporation) for months, which hugely reduces the amount of degassing that you need to do. You will only use say 0.5 ul of stock per drop. (reagents including oils stored in a tray in the glove box to reduce degassing time) 2. The oil offers a surprising amount of protection from oxidation, which may be helpful eg in harvesting. Perhaps by the O2-buffering effect of anoxic oil that Tristan mentioned, and by preventing an air/water interface where a skin forms and interferes with harvesting. 3. Microbatch can be automated - in parallel to vapor diffusion if desired And now as an aside, the advantages of oil for aerobic (oxic) work: It's amazing how often (aerobic) microbatch produces far superior crystals to V.D. for no obvious reason - it's well worth trying for both screening and optimization. If that was what was meant I apologize, but at least this will prevent someone else from getting the wrong Idea as I did. eab
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Hi Stephen, we crystallized several proteins in a glove box placed in a room with air condition. We checked the temperature all the times and found that is had been quite stable. For one protein that crystallized only at 4 deg C, we setup the crystallization plate within the glove box and put it into an gas-tight plexiglass box (built by our workshop). The box with the crystallization plate was then incubated at 4degC. Best, Guenter Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm.
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Thanks for all of the responses, there seems to be no real consensus, but I have discovered: 1 There are several small incubators supplied by - Molecular Dimensions, Revsci, Centeo 2 Good air conditioning may negate the need for an incubator. 3 A Glove box may not even be necessary depending on the samples sensitivity to oxygen. Certainly plenty for me to think about before I commit to buying a glove box. Yet again the bulletin board proves to be a great source of information for all things crystallographic! Best wishes, Steve Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Stephen Carr [stephen.c...@rc-harwell.ac.uk] Sent: 11 March 2015 10:17 To: ccp4bb Subject: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm.
[ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm.
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Dear Steve, There is a model of small incubator that fits through the large portal of a Belle Technology rigid glovebox - as an aside, that’s a good box for crystallisation and crystal manipulation since you can get the microscope-mounted side box with a dewar port for rapid and pretty efficient cryo-cooling. - I don’t work for Belle and don’t even know for sure if they are still in business.. Anyway…to your question...the incubator is constructed of grey plastic but holds temperature nicely - let me ring the guys back at Wake Forest Univ to see if they have it still around in the lab and I will send you the manufacturer and model number, or maybe they will see this here and post. Cheers -Conn Conn Mallett, PhD Director of Sales - SCX SBU Rigaku 9009 New Trails Drive The Woodlands, TX 77381 Office: +1 281 362 2300 ext. 126 Cell: +1 713 614 6891 FAX: +1 281 364 3628 Subscribe to Rigaku eNewsletters http://www.rigaku.com/subscribe . conn.mall...@rigaku.commailto:conn.mall...@rigaku.com www.rigaku.comhttp://www.rigaku.com On 11Mar, 2015, at 09:11, Martin Montgomery m...@mrc-mbu.cam.ac.ukmailto:m...@mrc-mbu.cam.ac.uk wrote: Dear Steve, You should not require an incubator if you have good, stable air con. We routinely grow crystals in plates stored at room temperature in wall mounted cabinets in our crystallisation lab. Another of the groups here have experience of growing crystals in an anaerobic box. Hopefully they will post more information. Regards MGM Martin G Montgomery ATP Synthase Group MRC Mitochondrial Biology Unit Wellcome Trust/MRC Building Cambridge Biomedical Campus Hills Road Cambridge Great Britain CB2 0XY www.mrc-mbu.cam.ac.ukhttp://www.mrc-mbu.cam.ac.uk/ On 11 Mar 2015, at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.ukmailto:stephen.c...@rc-harwell.ac.uk Stephen Carr wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.ukmailto:stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm.
Re: [ccp4bb] Off-topic - Crystallisation in anaerobic glove box
Dear Steve, You should not require an incubator if you have good, stable air con. We routinely grow crystals in plates stored at room temperature in wall mounted cabinets in our crystallisation lab. Another of the groups here have experience of growing crystals in an anaerobic box. Hopefully they will post more information. Regards MGM Martin G Montgomery ATP Synthase Group MRC Mitochondrial Biology Unit Wellcome Trust/MRC Building Cambridge Biomedical Campus Hills Road Cambridge Great Britain CB2 0XY www.mrc-mbu.cam.ac.uk On 11 Mar 2015, at 10:17, Stephen Carr stephen.c...@rc-harwell.ac.uk Stephen Carr wrote: Dear CCP4BBer's Apologies for the off-topic post, but the CCP4BB seems to be the best place to ask about crystallisation. I am looking to set up crystallisation in an anaerobic glove box and wondered how other people did this, specifically the crystallisation stage. My initial thoughts were to place a small crystallisation incubator inside the box, however the smallest I have come across so far (~27L) is still rather large. Has anyone come across smaller incubators? Alternatively are incubators even neccessary if the glove box is placed in a room with good air conditioning and stable temperature control? Any recommendations would be very helpful. Thanks in advance, Steve Carr Dr Stephen Carr Research Complex at Harwell (RCaH) Rutherford Appleton Laboratory Harwell Oxford Didcot Oxon OX11 0FA United Kingdom Email stephen.c...@rc-harwell.ac.uk tel 01235 567717 This email and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorized recipient of the addressee, please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to this email. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Research Complex at Harwell. There is no guarantee that this email or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. We use an electronic filing system. Please send electronic versions of documents, unless paper is specifically requested. This email may have a protective marking, for an explanation, please see: http://www.mrc.ac.uk/About/informationandstandards/documentmarking/index.htm.
