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Reservoir Solution (reservoir + solution)
Selected AbstractsA simple technique to convert sitting-drop vapor diffusion into hanging-drop vapor diffusion by solidifying the reservoir solution with agaroseJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 5 2009Tae Woong Whon A simple protocol to convert sitting-drop vapor-diffusion plating into a hanging-drop vapor-diffusion experiment in protein crystallization is reported. After making a sitting-drop plate, agarose solution was added to solidify the reservoir solution, and the plates were incubated upside down. Crystallization experiments with hen egg white lysozyme, thaumatin and glucose isomerase showed that the `upside-down sitting-drop' method could produce single crystals with all the benefits of the hanging-drop crystallization method. [source] A procedure for setting up high-throughput nanolitre crystallization experiments.ACTA CRYSTALLOGRAPHICA SECTION D, Issue 6 2005Crystallization workflow for initial screening, automated storage, imaging, optimization Crystallization trials at the Division of Structural Biology in Oxford are now almost exclusively carried out using a high-throughput workflow implemented in the Oxford Protein Production Facility. Initial crystallization screening is based on nanolitre-scale sitting-drop vapour-diffusion experiments (typically 100,nl of protein plus 100,nl of reservoir solution per droplet) which use standard crystallization screening kits and 96-well crystallization plates. For 294,K crystallization trials the barcoded crystallization plates are entered into an automated storage system with a fully integrated imaging system. These plates are imaged in accordance with a pre-programmed schedule and the resulting digital data for each droplet are harvested into a laboratory information-management system (LIMS), scored by crystal recognition software and displayed for user analysis via a web-based interface. Currently, storage for trials at 277,K is not automated and for imaging the crystallization plates are fed by hand into an imaging system from which the data enter the LIMS. The workflow includes two procedures for nanolitre-scale optimization of crystallization conditions: (i) a protocol for variation of pH, reservoir dilution and protein:reservoir ratio and (ii) an additive screen. Experience based on 592 crystallization projects is reported. [source] Crystallization and preliminary X-ray analysis of a recombinant Fab fragment in complex with 17,-oestradiolACTA CRYSTALLOGRAPHICA SECTION D, Issue 12 2000Urpo Lamminmäki The recombinant Fab fragment of the anti-17,-oestradiol antibody 57-2 has been a target for several protein-engineering experiments. A method for production, purification and crystallization of the Fab fragment alone (apo form) and in complex with the major female sex hormone 17,-oestradiol is reported here. Diffracting apo-form crystals were only obtained with microseeding; crystals of the Fab,steroid complex were produced by co-crystallization in the presence of oestradiol and cross-seeding with the apo-form crystals. The crystals were grown using vapour-diffusion methods with reservoir solutions containing 10,14% PEG 4000 or 8,12% PEG 8000 and Tris,HCl buffer at high pH (9.0,9.5). Both the apo and complex crystals belong to space group P212121 and diffract to 2.0,Å resolution. High-resolution X-ray data sets suitable for structure determination were collected from flash-cooled crystals using 25% glycerol as the cryoprotectant. [source] | ||||||||||