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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.


Related in: MedlinePlus

Dehydration of crystallization drops. Volumes of crystallization drops as a function of time. Each point represents the average diameter for eight crystallization droplets. Standard deviations of 1σ are typically below 15% of the calculated volumes (not shown).
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fig3: Dehydration of crystallization drops. Volumes of crystallization drops as a function of time. Each point represents the average diameter for eight crystallization droplets. Standard deviations of 1σ are typically below 15% of the calculated volumes (not shown).

Mentions: The rates of decrease in the calculated volumes of the droplets under the various vapor pressure conditions created by the solutions in the reservoirs are displayed in Fig. 3 ▶. The data clearly show that changing the salt concentration of the aqueous solution in the reservoir, and thereby the vapor pressure, dramatically changes the rate of decrease in the volumes of the droplets. The lower the salt concentration, the higher the vapor pressure, and the lower the rate of decrease in volume. With water in the reservoirs, it was effectively possible to completely prevent concentration of the droplets for periods of as long as two years, under which conditions crystals in the drops maintained their integrity and diffracting power. In some experiments, the trays were wrapped in parafilm, but this had no effect on the rate of evaporation (not shown).


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)

Dehydration of crystallization drops. Volumes of crystallization drops as a function of time. Each point represents the average diameter for eight crystallization droplets. Standard deviations of 1σ are typically below 15% of the calculated volumes (not shown).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: Dehydration of crystallization drops. Volumes of crystallization drops as a function of time. Each point represents the average diameter for eight crystallization droplets. Standard deviations of 1σ are typically below 15% of the calculated volumes (not shown).
Mentions: The rates of decrease in the calculated volumes of the droplets under the various vapor pressure conditions created by the solutions in the reservoirs are displayed in Fig. 3 ▶. The data clearly show that changing the salt concentration of the aqueous solution in the reservoir, and thereby the vapor pressure, dramatically changes the rate of decrease in the volumes of the droplets. The lower the salt concentration, the higher the vapor pressure, and the lower the rate of decrease in volume. With water in the reservoirs, it was effectively possible to completely prevent concentration of the droplets for periods of as long as two years, under which conditions crystals in the drops maintained their integrity and diffracting power. In some experiments, the trays were wrapped in parafilm, but this had no effect on the rate of evaporation (not shown).

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.


Related in: MedlinePlus