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Straightforward temperature screening
It has long been recognised that temperature should be utilized more effectively in the crystallization of bio-molecules.
With targets becoming ever more difficult to crystallize, introducing a simple and effective temperature screen to induce and optimise crystallization has never been more important.
The TG-40 opens the door to the routine sampling of temperature in everyday crystallography:
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Why screen temperature?
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The Science:
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Making temperature work for you - the three I s :
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Increasing crystallization space alongside precipitant, pH,
buffer etc. |
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| The ability to easily sample an incremental range of temperatures in a single location can yield both new crystal hits and improved crystal forms. | |
| Probe solubility - around 80% of proteins display a temperature dependence(1,2), as well as the phase behaviour of protein-detergent solutions.(3,4) |
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| Influence the kinetics of crystallization by altering diffusion and equilibrium rates.(5) |
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| Inducing crystal growth - increasing supersaturation: | |
| A controlled slow drop in temperature is a powerful strategy rarely employed in macromolecular crystallography. In lower salt systems this can fine tune the level of supersaturation and hence nucleation(6). | |
| Improving crystal quality - uncoupling nucleation from growth(7): | |
Protein crystal improvement – images courtesy of Andrew Bent. | |
| A temperature shift(6) or temperature cycling (combined with seeding)(8) during a crystallization can yield dramatic improvements in both size and quality. Temperature screening can also resolve twinning and alter the crystal space group.
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The Solution:
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Features of the TG-40:
- Accurately control and screen 5 different temperatures simultaneously - one set temperature per row.
- At 20ºC ambient the temperature range is 4ºC to 60ºC (accuracy is 1.0ºC).
- Portable operation- built-in battery.
- Disposable inserts (3µl sub well, 60µl main well) avoid cross-contamination.
- 40 wells (5x8) of standard SBS pitch for ease-of-use with automation.
- Simple, intuitive software for protocol set- up and recording.
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Click here for more information, product specs etc....
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References:
(1) Christopher et al. (1998) J. Cryst. Growth 191,820.
(2) Zhu et al. (2006) J. Struct. Biol. 154,297.
(3) Garavito & Picot (1991) J. Cryst. Growth 110,89.
(4) Lorber et al. (1991) J. Cryst. Growth 110,103.
(5) Lorber & Giegé (1992) J. Cryst. Growth 122,168.
(6) L. Lloyd Haire, in T.M. Bergfors(ed) Protein Crystallization (1st edition), I.U.L. 1999, pp. 65-68.
(7) Penkova et al. (2002) Acta Cryst. D58,1606.
(8) Budayova-Spano et al. (2007) Acta Cryst. D63, 339.
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