Tissue Culture 1 Cell/ Microplates 2 HTS- 3 Immunology/ HLA 4 Microbiology/ Bacteriology Purpose Beakers 5 Tubes/Multi-

Transcription

1 11 Cryo 5 Tubes/Multi 2 HTS

2 Technical Information 8 I 2 Vapour Diffusion Applications 8 I 4 Well CrystalQuick TM 8 I 4 CrystalDrop TM Lid 8 I 6 24 Well ComboPlate TM 8 I 7 CrystalBridge TM 8 I 7 Microbatch under Oil Applications 8 I 8 60 Well Terasaki Plate 8 I 8 72 Well Terasaki Plate 8 I 8 Well IMP@CT TM Plate 8 I Well IMP@CT TM Plate 8 I 9 11 Cryo 2 HTS 5 Tubes/Multi

3 2 HTS 5 Tubes/Multi Protein 11 Cryo An important method for the determination of protein structures is xray analysis of protein crystals. The determination of the threedimensional structure of proteins has contributed towards major advances in basic research, particularly in the fields of structural genomics and structurebased drug design. The most commonly used method for the crystallisation of proteins is vapour diffusion which comprises both the sitting drop and hanging drop methods (Fig. 1a and Fig. 1b). One drop of protein solution is mixed with one drop of reagent solution and incubated together with a larger volume of reagent solution in a sealed well. Concentration gradients between the sample drop and the reservoir solution are balanced out by diffusion, which induces the crystallisation process if the correct conditions have been selected. cover slide silicone grease VIEWseal TM sample drop reagent solution a) Hanging Drop b) Sitting Drop c) Microbatch Figure 1: methods a) Hanging Drop b) Sitting Drop c) Microbatch oil sample drop The microbatch method (Fig. 1c) in which the sample drop is covered with oil is also widely used, and in this technique the choice of oil determines the rate of diffusion of the water in the sample drop through the oil. Numerous factors affect the crystallisation of proteins. Since the optimal crystallisation conditions generally can t be predicted, a large number of attempts is often necessary in order to determine and optimise the appropriate conditions. Protein crystallisation therefore still represents a major bottleneck in structure analysis. The use of highthroughput technologies, such as pipetting robots and standardised microplates makes it possible to test a large number of crystallisation conditions in a short period of time and with relatively small amounts of protein. The CrystalStar product range from Greiner BioOne is a family of crystallisation plates and accessories designed specifically for highthroughput crystallisation. Format We place great value on the suitability of our protein crystallisation plates for use with automated systems. Therefore, with the exception of Terasaki plates, all crystallisation plates have a footprint conforming to the ANSI/SBS Standard. For further information please visit our website: /technical_information 8 2

4 ! Barcode Labelling Customerspecific barcode labelling is available on request for all crystallisation plates, with the exception of Terasaki plates. Material All Greiner BioOne protein crystallisation plates, with the exception of the plates (see below), are made from polystyrene. This is characterised by high clarity and excellent optical properties. Surface Treatment Surface treatment affects the polarity of the polystyrene surface of crystallisation wells. The surface is hydrophilised, which alters the wetting properties. Hydrophobic Plates Plates with a hydrophobic surface are particularly well suited for nanolitrecrystallisation of membrane proteins. The surface properties of hydrophobic plates efficiently counteract the spreading of detergentcontaining drops, respectively of drops with surfactant precipitants, such as MPD (Fig. 2). Moreover, the meniscus of the screening solution in the reservoir is substantially reduced, so that contaminations through creeping of the screening solution into the crystallisation well are avoided. Plates (low birefringence) plates are specifically designed for the use of polarised light. plates for sitting drop applications are made from polyolefin which is characterised by very low birefringence in comparison with polystyrene plates (Fig. ). Extreme transparency, high chemical resistance and low water absorption are further characteristics of plates. a) b) Figure 2: Comparison of (a) Plus (hydrophobic surface) and (b) standard. Images of 100 nl drops containing 50 mm noctylglucoside are courtesy of Karl Harlos, The Wellcome Trust Centre for Human Genetics, Oxford, UK. 1a) 2a) a) 1b) 2b) b) Figure : CrystalQuick TM plates in polarised light: (a) Standard versions with strong birefringence, (b) versions with lower birefringence. (1) CrystalQuick TM RW (2) CrystalQuick TM SW () CrystalQuick TM LP 2 HTS 5 Tubes/Multi Cat.No CrystalDrop Lid CrystalDrop Lid SW (Square Wells) SW (Square Wells) Plus SW (Square Wells) Plus SW (Square Wells) RW (Round Wells) RW (Round Wells) LP (Low Profile) LP (Low Profile) Plus LP (Low Profile) Number of sample wells Vapour Diffusion Microbatch Accessories Number of reservoirs Special features pregreased pregreased hydrophobic, hydrophobic hydrophobic Terasaki Plate Terasaki Plate IMP@CT Plate IMP@CT Plate IMP@CT Plate µclear film bottom 11 Cryo VIEWseal CrystalBridge Coverslip, 18 mm ø Coverslip, 22 mm ø Table 1: Overview of CrystalStar crystallisation plates and accessories. 1 glass, siliconised glass, siliconised 8

5 2 HTS Vapour Diffusion Applications Vapour Diffusion Applications Well Plates for Sitting Drop Applications 5 Tubes/Multi In cooperation with the Genomics Institute of the Novartis Research Foundation (GNF) in San Diego (USA), the Max Planck Institute (MPI) and the Protein Structure Factory (PSF) in Berlin, Greiner BioOne has developed a family of well crystallisation plates for sitting drop applications. Each of the reservoirs contains an elevated platform with one or three crystallisation wells. The plates are optimised for sealing with VIEWseal adhesive film. In combination with the CrystalDrop TM lid, simultaneous experiments are possible with the sitting drop and hanging drop method. The external dimensions and tolerances of the plates are suitable for automatised applications. All plates are available in an version for the use of polarised light. Plates with a hydrophobic surface can be found in the table under Plus plates. CrystalQuick TM SW (Square Wells Fig. 1, Fig. 2) With three crystallisation wells per reservoir, CrystalQuick TM SW makes it possible to test samples per plate. The bottom of the wells provides for good optical properties. The maximum volume of the crystallisation drops is 4 µl (US Patent No B2). Figure : Well profile, CrystalQuick TM RW Figure 4: Protein crystal in CrystalQuick TM RW, the image was kindly supplied by B. Blattmann, NCCR Structural, Switzerland CrystalQuick TM LP (Low Profile Fig. 5, Fig. 6) CrystalQuick TM LP (low profile) crystallisation plates are characterised by excellent optical properties. Crystal harvesting is made easier by the angled walls of the crystallisation wells. The low profile reduces space requirements for storage. 11 Cryo Figure 1: Well profile, CrystalQuick TM SW Figure 5: Well profile, CrystalQuick TM LP 0.50 mm Figure 2: of lysozyme in CrystalQuick TM SW CrystalQuick TM RW (Round Wells Fig., Fig. 4) With three round crystallisation wells per reservoir, CrystalQuick TM RW makes it possible to test samples per plate. The bottom of the crystallisation wells is concave. The maximum volume of the crystallisation drops is 1.9 µl. Figure 6: of lysozyme in CrystalQuick TM LP, RoboDesign International Inc., Carlsbad (USA) 8 4

6 Vapour Diffusion Applications Well CrystalQuick TM Well CrystalQuick TM Plus Lids and Sealers p. 12 I 2 ff. CrystalDrop TM lid for hanging drop applications p. 8 I 6 2 HTS plates for sitting drop applications with different well profiles and material properties CrystalQuick TM 5 Tubes/Multi Cat.No standard standard standard Well profile square (SW) square (SW) round (RW) round (RW) square (LP) square (LP) Well bottom Well per reservoir Max. well volume [µl] concave 1.9 concave Volume per reservoir [µl] Height [mm] (low profile) 20/ (low profile) 20/80 CrystalQuick TM Plus Cat.No Well profile Plus hydrophobic square (SW) Plus, hydrophobic square (SW) Plus hydrophobic square (LP) Well bottom Well per reservoir 1 11 Cryo Max. well volume [µl] Volume per reservoir [µl] Height [mm] (low profile) 20/80 8 5

7 2 HTS Vapour Diffusion Applications CrystalDrop TM Lid for Hanging Drop Applications CrystalDrop TM Lid Further Lids and Sealers p. 12 I 2 ff. CrystalQuick TM Plates p. 8 I 5 Lid for CrystalQuick TM microplates Enables individual sealing of the wells : Pregreased lids 5 Tubes/Multi The CrystalDrop lid was developed in cooperation with the Max Planck Institute (MPI) and the Protein Structure Factory (PSF) in Berlin specifically for use in highthroughput crystallisation. The 192 wells of CrystalDrop make it possible to place the sample drops at predefined positions. The printed grid facilitates the automated monitoring of the crystallisation drops. Channels around each of the positions enable the individual sealing of each well by the application of silicone grease. The combination of CrystalDrop and makes it possible to conduct up to sitting drop and 192 hanging drop experiments in parallel (a total of 480 crystallisation drops per plate). CrystalDrop is suitable for the application of crystallisation drops and the attachment to plates by robots. CrystalDrop is also available pregreased (Fig. 7). Figure 7: Pregreasing of CrystalDrop lid Cat.No Format CrystalDrop lid for plates well CrystalDrop lid for plates pregreased well Well per reservoir ø Well [mm] Length [mm] x width [mm] x height [mm] x 84.8 x x 84.8 x 6.0 5/80 5/40 11 Cryo 8 6

8 Vapour Diffusion Applications 24 Well and CrystalBridge TM Well CrystalBridge TM 2 HTS ComboPlate TM The ComboPlate TM was developed as universal platform for crystallisation in the 24 well format in cooperation with Hampton Research (Fig. 8). Clear polystyrene in combination with a, distortionfree bottom offers excellent optical properties. A tened, raised ring around each well reduces the risk of crosscontamination and makes it possible to seal the wells with silicone grease and coverslips (ø 18 mm) or VIEWseal sealer (Cat.No ). A slightly raised lid protects the coverslips and sealer during transportation and storage. Lids and Sealers p. 12 I 2 ff. Universal 24 well crystallisation plate! : Pregreased plates : Siliconised coverslips The is also available pregreased or in an version for use with polarised light now. As an accessory for the Greiner BioOne offers siliconised coverslips (round, ø 18 mm, Cat.No ) (Fig. 9). Siliconised coverslips for Linbro plates (round, ø 22 mm, Cat.No ) can also be found in our product line. 5 Tubes/Multi Figure 9: sealed with coverslips Figure 8: Well profile, 24 well ComboPlate TM CrystalBridge Sitting drop experiments are possible using the CrystalBridge inserts which fit exactly into the wells of the. The well with a concave bottom integrated into the CrystalBridge has a volume of 45 µl. If necessary, CrystalBridge inserts can be transferred to another well during the course of an experiment. Cat.No CrystalBridge Format Well bottom 24 well pregreased 24 well 24 well 1 well concave ø Well [mm] Max. well volume [µl] Lid Cryo Cat.No. 6/24 6/24 6/ round coverslips round coverslips ø [mm] Quantity per box/case siliconised glass /1000 siliconised glass /

9 2 HTS Microbatch under Oil Applications Microbatch under Oil Applications 60 Well and 72 Well Terasaki Plates Well / 72 Well Terasaki Plates Surfacetreated Terasaki Plates p. I 1 60 Well and 72 Well Terasaki Plates Terasaki plates are widely used for microbatch crystallisation. The crystallisation drop is localised centrally as a result of the conical well geometry, and the well bottom makes for optimal monitoring (Fig. 10). The rim of the Terasaki plates makes it possible to fill all of the wells with oil at the same time. As a result of the small external dimensions and the low profile of the Terasaki plates, the space required for storage is relatively small. Terasaki plates are supplied with a fitting lid. The plates are also supplied with surface treatment ( p. I 1). The treatment of the plates influences the sticking of the crystallisation drop to the bottom of the well. Figure 10: Well profile, Terasaki plate Cat.No Format Well profile Terasaki plate 60 well conical Terasaki plate 72 well conical Well bottom ø Well bottom [mm] Max. well volume [µl] Length [mm] x width [mm] x height [mm] Surface treatment 8. x 58 x 10 10/ x 58 x 10 10/ Cryo 5 Tubes/Multi 8 8

10 Well and 156 Well Plate Plates plates are microplates for highthroughput crystallisation, specially designed for microbatch applications under oil. The crystallisation plates are characterised by conical wells with a bottom. The conical form of the wells has the effect of centrally localising the crystallisation drops, even when small sample volumes are used. The well bottom makes for optimal monitoring of the crystallisation samples. An appropriate lid is available. Well IMP@CT Plate (Fig. 11) The new well IMP@CT plate was developed by Greiner BioOne in cooperation with Hampton Research (USA) and Allan D Arcy. The double rim of the plates means that the wells can be filled separately or together with oil. Filling the double rim of the IMP@CT plates with an aqueous gel makes it possible to control water evaporation from the crystallisation drops. The well IMP@CT plate is also available as a black µclear version with a film bottom. This plate is charcterised by extremely low light scattering between individual wells and by a low birefringence. Microbatch under Oil Applications 156 Well IMP@CT Plate (Fig. 12) The new 156 well IMP@CT plate was developed in cooperation with the HauptmanWoodward Medical Research Institute in Buffalo (USA) for use in automated highthroughput systems. The optimised well geometry means that even crystallisation drops with a very small volume are localised at the centre of the wells. Smooth, well bottoms provide optimum conditions for monitoring and evaluating crystallisation samples. The 156 well IMP@CT plate is suitable for use in polarised light. a) b) Figure 12a: Well profile, 156 Well IMP@CT plate Figure 12b: of lysozyme in the 156 well IMP@CT plate, with the kind approval of the HauptmanWoodward Medical Research Institute, Buffalo (USA). 2 HTS 5 Tubes/Multi Figure 11: Well profile, Well IMP@CT plate 67 0 Well / 156 Well IMP@CT Plate Lids and Sealers p. 12 I 2 ff. Plate for crystallisation under oil Cat.No Cryo IMP@CT IMP@CT IMP@CT Format Max. well volume [µl] clear well 8.0 black, µclear bottom well 8.0 clear, 156 well 10.1 ø Well bottom [mm] Height [mm] /60 8 9