Re: [ccp4bb] Membrane Protein Crystallization Plates
Yuri I know this isn't quite what you're asking, but it's helpful to use the COC (UV permeable) version of plates if you're crystallizing samples that contain detergent. It's just that the drops tend to spread very thinly if you use the normal polystyrene PS plates. The COC is more hydrophobic. (On the other hand we prefer PS plates for normal proteins with no detergent.) Patrick On 25 February 2012 20:35, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello Everyone, I am considering the purchase of crystallization plates for membrane proteins. I would love to hear what some of the community thinks or has experienced with these. Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) So fire away. Is it worth it? Any succes stories? Bad experiences? I appreciate the input Best, Yuri -- 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] Membrane Protein Crystallization Plates
Hi i do not want to get into trouble by going against any products The plates what you are talking about as soon it came out we tested. I did not look back to see if there was a monoolein:cholestrol coated plate. The ones i used were for sure monoolein coated ones. That tells it all.The rest of the plates still unused. I set up plates with bacteriorhodopsin even that did not work in our hands (the bacteriorhodopsin set up otherwise would work very well.) By the way it did not work for me did not mean it is bad. And the plates were in plasticware made it highly birifringent to document the results. But the plates Patrick says here i am not sure about. If Patrick want to to tell me a little bit more about i would like to hear that. I would set up plates the normal way. I am really not convinced mesophase will form by just adding protein-detergent to a previously poured monoolein. anyway i am also interested to hear experts views of the method adapted in this product. There is a poster that they shown at their website shows bacteriorhodopsin and another membrane protein crystallized using these plates. These are my personal opinion. Pius On Sat, Feb 25, 2012 at 3:35 PM, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello Everyone, I am considering the purchase of crystallization plates for membrane proteins. I would love to hear what some of the community thinks or has experienced with these. Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) So fire away. Is it worth it? Any succes stories? Bad experiences? I appreciate the input Best, Yuri -- Pius S Padayatti,PhD, Phone: 216-658-4528
Re: [ccp4bb] Membrane Protein Crystallization Plates
Yuri, Did you mean plates for setting up Lipidic Mesophases? If so, here is a listing of products I have used in the past. I highly endorse the plates from Molecular dimensions, particularly the plastic laminex plates (MD11-51-100 + MD11-54). They do have the dis-advantage of drying out after a 2-month or so period, but they make the job of harvesting significantly easier than dealing with the glass equivalents (so much so that I don't ever plan on using the glass bases and covers again). *Molecular Dimensions:* Using the MD11-51-100 plastic bases combined with the MD11-54 covers results in a cost of $9.20 per plate. Laminex Plastic 100 um base (MD11-51-100) - I've used these to solve multiple structures during my post-doc at NIH. They're UV transparent for doing UV microscopy and detection of your crystals. Cost: $66 USD for a pack of 10. Laminex Plastic 200 um base (MD11-50) - Has a thicker 200 um layer for inputting larger amounts of mesophase and precipitant. Cost: $66 USD for a pack of 10. Laminex Film Cover (MD11-54) - Used to cover the top of the sandwich plate. Also UV transparent. Cost: $26 USD per pack of 10. Laminex Glass 100 um base (MD11-50-100) - Glass base with 100 um spacer. Cost: $66 USD per pack of 10. Laminex Glass 200 um base (MD11-50) - Glass base with 200 um spacer. Cost: $66 USD per pack of 10. Glass Covers (MD11-52) - Glass for forming sandwich plates. Cost: $40 per pack of 10. *Hampton Research:* Plastic UV Transparent LCP Setup Kit (HR3-186) - Set of 20 plastic plates in a slightly different form factor than the Mol Dim. plastic plates. Cost: $483 USD per pack of 20. This is $24.15 per plate. Glass UV Transparent LCP Setup Kit (HR3-151) - Siliconized glass bases and glass tops. Cost: $368 USD per pack of 20 ($18.40 per plate). On Sat, Feb 25, 2012 at 12:35 PM, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello Everyone, I am considering the purchase of crystallization plates for membrane proteins. I would love to hear what some of the community thinks or has experienced with these. Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) So fire away. Is it worth it? Any succes stories? Bad experiences? I appreciate the input Best, Yuri -- Jim Fairman, Ph D. Crystal Core Leader I Emerald BioStructures http://www.emeraldbiostructures.com/ Tel: 206-780-8914 Cell: 240-479-6575 E-mail: fairman@gmail.com jfair...@embios.com
Re: [ccp4bb] Membrane Protein Crystallization Plates
QuoteParticulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) Since the person who asked this question here forget about it alltogether to write something back here is what he was asking about (i think) Anybody heard about the nextal plates see the link http://www.qiagen.com/products/nextalcubicphaseproducts.aspx It uses a different method of mixing lipids other than using the syringe method. They have lipid layered on the bottom of the well and protein samples are pipetted onto the monoolein.and later on precipitant. Plastic based ones are really bad idea Not everyone have an access to UV even if UV screening is used not a full-proof method to detect membrane protein micro crystals I would definitely think glass based plates are the way to go. On Sat, Feb 25, 2012 at 3:35 PM, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello Everyone, I am considering the purchase of crystallization plates for membrane proteins. I would love to hear what some of the community thinks or has experienced with these. Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) So fire away. Is it worth it? Any succes stories? Bad experiences? I appreciate the input Best, Yuri -- Pius S Padayatti,PhD, Phone: 216-658-4528
Re: [ccp4bb] Membrane Protein Crystallization Plates
Thanks for all the replies. I will try a couple of different plates/set-ups. My favorite will be the one that gives me a crystal ;)
Re: [ccp4bb] Membrane Protein Crystallization Plates
Hi Yuri, i strongly suggest these plates sold through Hampton research Paul Marienfeld GmbH plates for your use. here is a link , best plates in the market. link: http://hamptonresearch.com/product_detail.aspx?cid=10sid=182pid=611 Plates comes with extra cover slips in addition to single glass cover for four wells at a time whcih let you set these plates manually if you like it that way. Good for automations as well. only problem: since the galss is siliconized you can not write anything on it (may be there is some industry grade marker might work.) But there is enough place on side to stick a printed label or bar code for your'automation for keeping records hope this is what you wanted to know. if you are money crunched and want to make some plates on your own. buy regular micoscope slides, some two sided tapes (3M) very thin and a tool that sold for cutting out small pieces of tissue samples. it looks like a pen with sharp punch enough to make a hole in a 3M tape make a 27 hole plate in a standard microslide. cheers pius On Sat, Feb 25, 2012 at 3:35 PM, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello Everyone, I am considering the purchase of crystallization plates for membrane proteins. I would love to hear what some of the community thinks or has experienced with these. Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it should not matter) So fire away. Is it worth it? Any succes stories? Bad experiences? I appreciate the input Best, Yuri -- Pius S Padayatti,PhD, Phone: 216-658-4528
