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Control of the rate of evaporation in protein crystallization by the 'microbatch under oil' method.

Brumshtein B, Greenblatt HM, Futerman AH, Silman I, Sussman JL - J Appl Crystallogr (2008)

Bottom Line: Microbatch crystallization under oil is a powerful procedure for obtaining protein crystals.Using this method, aqueous protein solutions are dispensed under liquid oil, and water evaporates through the layer of oil, with a concomitant increase in the concentrations of both protein and precipitant until the nucleation point is reached.A technique is presented for regulating the rate of water evaporation, which permits fine tuning of the crystallization conditions as well as preventing complete desiccation of the drops in the microbatch crystallization trays.

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ABSTRACT
Microbatch crystallization under oil is a powerful procedure for obtaining protein crystals. Using this method, aqueous protein solutions are dispensed under liquid oil, and water evaporates through the layer of oil, with a concomitant increase in the concentrations of both protein and precipitant until the nucleation point is reached. A technique is presented for regulating the rate of water evaporation, which permits fine tuning of the crystallization conditions as well as preventing complete desiccation of the drops in the microbatch crystallization trays.

No MeSH data available.


Image of a crystallization droplet under oil in a well in the crystallization plate. The measured diameter of the droplet is indicated by the double-headed arrow.
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fig2: Image of a crystallization droplet under oil in a well in the crystallization plate. The measured diameter of the droplet is indicated by the double-headed arrow.

Mentions: In each plate, 48 droplets were dispensed. In order to follow evaporation as a function of time, eight droplets at identical locations in all the trays were selected for monitoring. Their diameter was measured using an Olympus microscope equipped with an XR12 lens, which yielded ×90 magnification. The horizontal diameter of the droplet was measured using the microscope scale bar (Fig. 2 ▶), making the assumption that the droplets are spherical. This assumption was shown to be valid by measuring their initial diameters, calculating the volume by use of the formula V = (4/3)πr 3, and comparing this calculated volume with the volume dispensed. Measurements of droplet diameters were made daily for 14 d. The plates were maintained at 295 K and at an external humidity of 65–75%.


Control of the rate of evaporation in protein crystallization by the 'microbatch under oil' method.

Brumshtein B, Greenblatt HM, Futerman AH, Silman I, Sussman JL - J Appl Crystallogr (2008)

Image of a crystallization droplet under oil in a well in the crystallization plate. The measured diameter of the droplet is indicated by the double-headed arrow.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2553557&req=5

fig2: Image of a crystallization droplet under oil in a well in the crystallization plate. The measured diameter of the droplet is indicated by the double-headed arrow.
Mentions: In each plate, 48 droplets were dispensed. In order to follow evaporation as a function of time, eight droplets at identical locations in all the trays were selected for monitoring. Their diameter was measured using an Olympus microscope equipped with an XR12 lens, which yielded ×90 magnification. The horizontal diameter of the droplet was measured using the microscope scale bar (Fig. 2 ▶), making the assumption that the droplets are spherical. This assumption was shown to be valid by measuring their initial diameters, calculating the volume by use of the formula V = (4/3)πr 3, and comparing this calculated volume with the volume dispensed. Measurements of droplet diameters were made daily for 14 d. The plates were maintained at 295 K and at an external humidity of 65–75%.

Bottom Line: Microbatch crystallization under oil is a powerful procedure for obtaining protein crystals.Using this method, aqueous protein solutions are dispensed under liquid oil, and water evaporates through the layer of oil, with a concomitant increase in the concentrations of both protein and precipitant until the nucleation point is reached.A technique is presented for regulating the rate of water evaporation, which permits fine tuning of the crystallization conditions as well as preventing complete desiccation of the drops in the microbatch crystallization trays.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Microbatch crystallization under oil is a powerful procedure for obtaining protein crystals. Using this method, aqueous protein solutions are dispensed under liquid oil, and water evaporates through the layer of oil, with a concomitant increase in the concentrations of both protein and precipitant until the nucleation point is reached. A technique is presented for regulating the rate of water evaporation, which permits fine tuning of the crystallization conditions as well as preventing complete desiccation of the drops in the microbatch crystallization trays.

No MeSH data available.