Re: [ccp4bb] optimization of dumbbell like crystal
Additives screening. YB 2011-02-21 yybbll 发件人： Bosch, Juergen 发送时间： 2011-02-21 03:46:12 收件人： CCP4BB 抄送： 主题： Re: [ccp4bb] optimization of dumbbell like crystal Seeding ? changing from sitting to hanging drop or under oil ? play with temperature ? Jürgen On Feb 20, 2011, at 9:29 PM, Harvey Rodriguez wrote: Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey dumbbell crystals.jpg .. Jürgen Bosch Johns Hopkins Bloomberg School of Public Health Department of Biochemistry Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Phone: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-3655 http://web.mac.com/bosch_lab/
Re: [ccp4bb] optimization of dumbbell like crystal
Dear Harvey, Microseeding and adding 1-5% glycerol to the drops helped me when I obtained crystals like these. Good Luck! Bryan From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Harvey Rodriguez Sent: Sunday, February 20, 2011 9:29 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] optimization of dumbbell like crystal Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey -- Confidentiality Notice: This message is private and may contain confidential and proprietary information. If you have received this message in error, please notify us and remove it from your system and note that you must not copy, distribute or take any action in reliance on it. Any unauthorized use or disclosure of the contents of this message is not permitted and may be unlawful.
Re: [ccp4bb] optimization of dumbbell like crystal
Dear Harvey, I had very good results with low melt agarose and the detergent screen (e.g. Hampton Research). In both cases, the tendency for cluster formation as well as the diffraction power could be improved. Anja Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey Anja Pomowski Universität Freiburg Institut f. Organ. Chemie und Biochemie AK Prof. Einsle Albertstr. 21 79104 Freiburg Tel. 0761 2036088 Fax. 0761 2036161
Re: [ccp4bb] optimization of dumbbell like crystal
Seeding ? changing from sitting to hanging drop or under oil ? play with temperature ? Jürgen On Feb 20, 2011, at 9:29 PM, Harvey Rodriguez wrote: Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey dumbbell crystals.jpg .. Jürgen Bosch Johns Hopkins Bloomberg School of Public Health Department of Biochemistry Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Phone: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-3655 http://web.mac.com/bosch_lab/
Re: [ccp4bb] optimization of dumbbell like crystal
Dear Harvey Based on the experimental history you provided I would try two quick things in order to obtain additional dimensions for your condition. (1) set up your best consensus condition against an additive screen (e.g. From Hampton). (2) I would repeat your last pH vs precipitant grid against different starting protein concentrations. You can conveniently play with different drop ratios to screen for that. Best regards Savvas On 21 Feb 2011, at 03:29, Harvey Rodriguez h.rodriguez.x...@gmail.com wrote: Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey dumbbell crystals.jpg
Re: [ccp4bb] optimization of dumbbell like crystal
Dear harvey Consider your purification and preparation protocols - is the sample pure and homogenous? Poul On 21/02/2011, at 03.29, Harvey Rodriguez h.rodriguez.x...@gmail.com wrote: Dear CCP4BBers, Recently I got a crystal which appeared in the condition 2.1M DL-Malic Acid, pH 7.0. The crystal looks like a dumbbell but was composed of a cluster of very thin needles as shown in the picture. The crystal can be repeated but the diffraction is poor. I have tried the grid optimization by pH and salt concentration gradients, however there was no improvement in either the shape or the quality of the crystal. Does anyone have some experiences in optimizing this kind of crystals? Any suggestion is appreciated! Harvey dumbbell crystals.jpg
