Re: [ccp4bb] Improving microcrystals
I've had some success in the past following Terese Bergfors' advice: http://xray.bmc.uu.se/terese/tutorial3.html However, the last time I dealt with spherulites I just tuned the pH and it worked like a charm. Good luck, Jon 2012/8/25 RHYS GRINTER r.grinte...@research.gla.ac.uk Hi Samuel, I've has good success going from sphereulites to crystals using an additive screen (the 96 condition Hampton one is good) with the conditions giving the spherulites. Just watch for salt crystals as you'll be adding some compounds that might cause your Ca ions to form insoluble CaSO4. http://hamptonresearch.com/product_detail.aspx?cid=1sid=36pid=27 Cheers, Rhys From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] On Behalf Of Samuel Johnson [samueljohnson...@yahoo.in] Sent: 25 August 2012 02:11 To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Improving microcrystals Hi everyone, I have been working on a protein for the past year. After a number of trials at crystallizing the protein i have identified conditions for getting spherulites/micro-crystaline material under micro batch method. I have confirmed that the crystalline material is protein, by using Izit-dye test. The condition is 50mM CaCl2, Mes pH 6.5 and 40% PEG 400. I will be happy to get suggestions on improving conditions to obtain single crystals. I have already tried varying a number of parameters like salt, precipitant concentration and buffer pH but that didn't help. Thanks. -- Jon Agirre, PhD Unit of Biophysics (CSIC-UPV/EHU) http://www.ehu.es/jon.agirre http://sourceforge.net/projects/projectrecon/ +34656756888
Re: [ccp4bb] Improving microcrystals
Samuel Clearly you should try the rMMS random microseeding approach where you add a seed stock made from the spherulites to a random screen. See refs: Acta Crystallographica section D63 (2007), 550–554. On-line at http://scripts.iucr.org/cgi-bin/paper?S0907444907007652 Cryst. Growth Des., 2011, 11 (8), pp 3432–3441. On-line at http://pubs.acs.org/doi/abs/10.1021/cg2001442. http://www.douglas.co.uk/SER-CAT09_1.html On 25 August 2012 02:11, Samuel Johnson samueljohnson...@yahoo.in wrote: Hi everyone, I have been working on a protein for the past year. After a number of trials at crystallizing the protein i have identified conditions for getting spherulites/micro-crystaline material under micro batch method. I have confirmed that the crystalline material is protein, by using Izit-dye test. The condition is 50mM CaCl2, Mes pH 6.5 and 40% PEG 400. I will be happy to get suggestions on improving conditions to obtain single crystals. I have already tried varying a number of parameters like salt, precipitant concentration and buffer pH but that didn't help. Thanks. -- 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
[ccp4bb] Improving microcrystals
Hi everyone, I have been working on a protein for the past year. After a number of trials at crystallizing the protein i have identified conditions for getting spherulites/micro-crystaline material under micro batch method. I have confirmed that the crystalline material is protein, by using Izit-dye test. The condition is 50mM CaCl2, Mes pH 6.5 and 40% PEG 400. I will be happy to get suggestions on improving conditions to obtain single crystals. I have already tried varying a number of parameters like salt, precipitant concentration and buffer pH but that didn't help. Thanks.
