Hampton Santovac Cryo Oil替代HR2-643 Parabar10312

Hampton Santovac Cryo Oil替代HR2-643 Parabar10312

APPLICATIONS

  • Oil based cryoprotectant

应用
油基冷冻保护剂

FEATURES

  • Low viscosity
  • Low surface tension
  • Cryoprotectant for soluble and membrane protein crystals
  • Not miscible with most solvents
  • Non toxic, non penetrating cryoprotectant

特征
低粘度
低表面张力
可溶性蛋白和膜蛋白晶体的冷冻保护剂
与大多数溶剂不混溶
无毒、非渗透性冷冻保护剂

DESCRIPTION

Used during cryocrystallography to displace and reduce the amount of water (mother liquor, reagent) on the crystal after the crystal is mounted in a cryoloop. Coating the crystal with oil can minimize evaporation from the crystal and reduce exposure and slow diffusion of air (oxygen) to the crystal.

Santovac Cryo Oil offers low viscosity, incredibly low vapor pressure, low surface tension, excellent chemical inertness and excellent thermal stability.

Dip the mounted crystal into the oil to displace surface water. Or add a small amount of Santovac to the crystallization drop before mounting the crystal. After adding the Santovac, mount the crystal using a Mounted Cryoloop. Withdraw the mounted crystal from the drop and the Santovac will coat the mounted crystal. Cryogenically cool the mounted crystal.

Santovac (Santovac 5) is a five ring polyphenyl ether (PPE) with very low vapor pressure and low reactivity. The oil is highly resistant to oxidation. The only elements in the pure polyphenyl ether are oxygen, hydrogen and carbon.

The fluid is chemically stable, non-corrosive, safe and non-toxic at normal operating temperatures.

Santovac is not miscible with water, paraffin oil, silicon oil, alkanes, or Parabar 10312 (Paratone). Santovac is miscible with acetone. Shelf life 5 years.

Santovac® is a trademark of Santovac® Fluids.

描述
在冷冻晶体学期间用于在晶体安装在冷冻环中后置换和减少晶体上的水(母液、试剂)的量。用油涂覆晶体可以最大限度地减少晶体的蒸发,并减少空气(氧气)对晶体的暴露和缓慢扩散。

Santovac Cryo Oil 具有低粘度、极低的蒸气压、低表面张力、出色的化学惰性和出色的热稳定性。

将镶嵌好的晶体浸入油中以置换地表水。或在安装晶体之前向结晶滴中添加少量 Santovac。添加 Santovac 后,使用 Mounted Cryoloop 安装晶体。从液滴中取出已安装的晶体,Santovac 将涂覆已安装的晶体。低温冷却安装的晶体。

Santovac (Santovac 5) 是一种具有极低蒸气压和低反应性的五环聚苯醚 (PPE)。该油具有很强的抗氧化性。纯聚苯醚中唯一的元素是氧、氢和碳。

该流体在正常工作温度下化学性质稳定、无腐蚀性、安全且无毒。

Santovac 与水、石蜡油、硅油、烷烃或 Parabar 10312 (Paratone) 不混溶。 Santovac 可与丙酮混溶。保质期5年。

Santovac® 是 Santovac® Fluids 的商标。

HR2-861 Santovac Cryo Oil 1 mL

RELATED ITEM(S)

REFERENCES

1. Structure of the ‘open’ form of Aspergillus nidulans 3-dehydroquinate synthase at 1.7 Å resolution from crystal grown following enzyme turnover. C.E. Nichols, A.R. Hawkins and D.K. Stammersa. Acta Crystallographic Section D, Volume 60, Part 5, Pages 971-973, May 2004.

2. Crystal Structure of the Caspase Activator Human Granzyme B, a Proteinase Highly Specific for an Asp-P1 Residue. E. Estebanez-Perpina et al. Biol. Chem., Vol. 381, pp. 1203-1214, December 2000.

Hampton HR2-861 Santovac Cryo Oil 油基防冻剂

Hampton HR2-861 Santovac Cryo Oil 油基防冻剂

Hampton research蛋白结晶试剂代理–上海金畔生物
代理优势:货期3-4周。
欢迎访问Hampton research官网或者咨询我们获取更多产品信息。

 Optimize Reagents Optimize – Cryoprotectants Santovac Cryo Oil

APPLICATIONS

  • Oil based cryoprotectant

应用
油基防冻剂

FEATURES

  • Low viscosity
  • Low surface tension
  • Cryoprotectant for soluble and membrane protein crystals
  • Not miscible with most solvents
  • Non toxic, non penetrating cryoprotectant

特征
低粘度
低表面张力
可溶性和膜蛋白晶体的冷冻保护剂
不与大多数溶剂混溶
无毒、非渗透性冷冻保护剂

DESCRIPTION

Used during cryocrystallography to displace and reduce the amount of water (mother liquor, reagent) on the crystal after the crystal is mounted in a cryoloop. Coating the crystal with oil can minimize evaporation from the crystal and reduce exposure and slow diffusion of air (oxygen) to the crystal.

Santovac Cryo Oil offers low viscosity, incredibly low vapor pressure, low surface tension, excellent chemical inertness and excellent thermal stability.

Dip the mounted crystal into the oil to displace surface water. Or add a small amount of Santovac to the crystallization drop before mounting the crystal. After adding the Santovac, mount the crystal using a Mounted Cryoloop. Withdraw the mounted crystal from the drop and the Santovac will coat the mounted crystal. Cryogenically cool the mounted crystal.

Santovac (Santovac 5) is a five ring polyphenyl ether (PPE) with very low vapor pressure and low reactivity. The oil is highly resistant to oxidation. The only elements in the pure polyphenyl ether are oxygen, hydrogen and carbon.

The fluid is chemically stable, non-corrosive, safe and non-toxic at normal operating temperatures.

Santovac is not miscible with water, paraffin oil, silicon oil, alkanes, or Parabar 10312 (Paratone). Santovac is miscible with acetone. Shelf life 5 years.

Santovac® is a trademark of Santovac® Fluids.

HR2-861  Santovac Cryo Oil 1 mL

描述
在晶体安装在低温回路中后,在低温晶体学期间用于置换和减少晶体上的水(母液、试剂)量。用油涂抹晶体可以最大限度地减少晶体的蒸发并减少空气(氧气)向晶体的暴露和缓慢扩散。

Santovac Cryo Oil 具有低粘度、极低的蒸气压、低表面张力、出色的化学惰性和出色的热稳定性。

将安装的晶体浸入油中以置换表面水。或者在安装晶体之前将少量的 Santovac 添加到结晶液滴中。添加 Santovac 后,使用 Mounted Cryoloop 安装晶体。从水滴中取出已镶嵌的水晶,Santovac 将覆盖已镶嵌的水晶。低温冷却安装的晶体。

Santovac (Santovac 5) 是一种五环聚苯醚 (PPE),具有极低的蒸气压和低反应性。该油具有很强的抗氧化性。纯聚苯醚中唯一的元素是氧、氢和碳。

该流体在正常工作温度下化学性质稳定、无腐蚀性、安全无毒。

Santovac 与水、石蜡油、硅油、烷烃或 Parabar 10312 (Paratone) 不混溶。 Santovac 可与丙酮混溶。保质期5年。

Santovac® 是 Santovac® Fluids 的商标。

HR2-861 Santovac 冷冻油 1 毫升

RELATED ITEM(S) 相关项目
CryoPro

Hampton HR2-861 Santovac Cryo Oil 油基防冻剂

REFERENCES  参考文献
1. Structure of the ‘open’ form of Aspergillus nidulans 3-dehydroquinate synthase at 1.7 Å resolution from crystal grown following enzyme turnover. C.E. Nichols, A.R. Hawkins and D.K. Stammersa. Acta Crystallographic Section D, Volume 60, Part 5, Pages 971-973, May 2004.

2. Crystal Structure of the Caspase Activator Human Granzyme B, a Proteinase Highly Specific for an Asp-P1 Residue. E. Estebanez-Perpina et al. Biol. Chem., Vol. 381, pp. 1203-1214, December 2000.

使用盐作为结晶试剂

Cryosalts   使用盐作为结晶试剂

Using salt as a crystallization reagent?

Many salts are cryosalts, including malonate, formate, citrate, tartrate, acetate, Tacsimate and other organic acids, ammonium sulfate (>3.5 M), lithium sulfate, lithium chloride and other alkylammonium salts. For cryo try increasing your salt concentration by 20%. (Cryosalts: suppression of ice formation in macromolecular crystallography, K. A. Rubinson et al, Acta Cryst. (2000). D56, 996-1001 doi:10.1107/S0907444900007587.

冰晶盐
使用盐作为结晶试剂?

很多盐是冰晶盐,包括丙二酸盐、甲酸盐、柠檬酸盐、酒石酸盐、乙酸盐、Tacsimate和其他有机酸、硫酸铵(>3.5 M)、硫酸锂、氯化锂和其他烷基铵盐。 对于冷冻,尝试将盐浓度提高 20%。 (冷冻盐:大分子晶体学中冰形成的抑制,K. A. Rubinson 等人,Acta Cryst. (2000). D56, 996-1001 doi:10.1107/S0907444900007587。

更换CryoLoop环的方法与建议

更换CryoLoop环的方法与建议

Replacing a Mounted CryoLoop in a CrystalCap 更换 CrystalCap 中已安装的 CryoLoop

Removal 
With one hand, using pliers, grasp the Mounted CryoLoop. Using your the other hand, hold the CrystalCap between your thumb and finger(s). With gentle authority twist the Mounted CryoLoop and Cap in opposing directions and pull the Mounted CryoLoop from the CrystalCap.

Replacement 
Select a Mounted CryoLoop with the desired CryoLoop size. Notice the notches on the Mounted CryoLoop. A standard length Mounted CryoLoop is created by snapping the MicroTube at the second notch from the bottom. Remove the end using your fingers or diagonal cutter. Apply a drop of Super Glue to the top opening of the CrystalCap. Insert the Mounted CryoLoop into this opening and push until the Mounted CryoLoop is stopped by the bottom of the CrystalCap. Epoxy may be used instead of Super Glue for a more permanent fixation of the Mounted CryoLoop to the CrystalCap.

移动
用一只手用钳子抓住已安装的 CryoLoop。 用另一只手将 CrystalCap 夹在拇指和其他手指之间。 以温和的权威将已安装的 CryoLoop 和 Cap 朝相反方向扭转,然后将已安装的 CryoLoop 从 CrystalCap 中拉出。

替代品
选择具有所需 CryoLoop 尺寸的已安装 CryoLoop。 注意已安装的 CryoLoop 上的槽口。 标准长度的已安装 CryoLoop 是通过在从底部开始的第二个凹口处卡入 MicroTube 来创建的。 用手指或斜切刀去除末端。 在 CrystalCap 的顶部开口处涂抹一滴强力胶。 将已安装的 CryoLoop 插入此开口并推动,直到已安装的 CryoLoop 被 CrystalCap 的底部挡住。 可以使用环氧树脂代替强力胶,以便将已安装的 CryoLoop 更永久地固定在 CrystalCap 上。

在普通结晶试剂添加硫酸铵和盐

Cryo for Ammonium sulfate and salt based crystallization reagents   用于硫酸铵和盐基结晶试剂的 Cryo

Increase the concentration of ammonium sulfate (cryosalt) (or primary salt acting as a precipitant) in increments of 10% in the presence of 5 to 10% v/v Glycerol.

Evaluate 25-30% w/v glucose, or trehalose or sucrose as cryoprotectants for crystals grown in Ammonium sulfate. Acta Cryst. (2002). D58, 1664-1669. Crystallization of RNA/protein complexes. M. Garber, G. Gongadze, V. Meshcheryakov, O. Nikonov, A. Nikulin, A. Perederina, W. Piendl, A. Serganov and S. Tishchenko.

15 to 30% v/v Ethylene glycol, DMSO, or Glycerol.

Try 1 – 2 M Sodium malonate as a cryoprotectant. Acta Cryst. (2003). D59, 2356-2358. Malonate: a versatile cryoprotectant and stabilizing solution for salt-grown macromolecular crystals. T. Holyoak, T. D. Fenn, M. A. Wilson, A. G. Moulin, D. Ringe and G. A. Petsko.

Place a drop of 75% v/v Paratone-N, 25% v/v Paraffin Oil next to the drop containing the crystal. Remove the crystal from the drop using a CryoLoop and dip the crystal into the oil. Keeping the CryoLoop and crystal immersed, gently move the CryoLoop containing the crystal from the oil into the reagent, back into the oil, back into the reagent, repeating this several times to remove some of the aqueous (reagent) layer from the crystal surface. Cryo cool the crystal.

在 5 至 10% v/v 甘油存在的情况下,以 10% 的增量增加硫酸铵(冰晶盐)(或用作沉淀剂的初级盐)的浓度。

评估 25-30% w/v 葡萄糖、海藻糖或蔗糖作为在硫酸铵中生长的晶体的冷冻保护剂。晶体学报。 (2002 年)。 D58,1664-1669。 RNA/蛋白质复合物的结晶。 M. Garber、G. Gongadze、V. Meshcheryakov、O. Nikonov、A. Nikulin、A. Perederina、W. Piendl、A. Serganov 和 S. Tishchenko。

15 至 30% v/v 乙二醇、DMSO 或甘油。

尝试 1 – 2 M 丙二酸钠作为冷冻保护剂。晶体学报。 (2003 年)。 D59,2356-2358。丙二酸盐:一种用于盐生大分子晶体的多功能冷冻保护剂和稳定溶液。 T. Holyoak、T. D. Fenn、M. A. Wilson、A. G. Moulin、D. Ringe 和 G. A. Petsko。

在含有晶体的液滴旁边放置一滴 75% v/v Paratone-N、25% v/v 石蜡油。使用 CryoLoop 从液滴中取出晶体并将晶体浸入油中。保持 CryoLoop 和晶体浸没,轻轻地将含有晶体的 CryoLoop 从油中移入试剂中,再移回油中,再移回试剂中,重复几次以去除晶体表面的一些水(试剂)层。低温冷却晶体。

Cleaning a CryoLoop   CryoLoop环的清洗方法

Cleaning a CryoLoop   CryoLoop环的清洗方法

Soak the CryoLoop in a 0.5% detergent solution in deionized water for 15 minutes. Gently rinse the CryoLoop in deionized water. Allow the CryoLoop to air dry.

If more rigorous cleaning is desired or one does not wish to risk the presence of trace amounts of residual detergent, soak the CryoLoop in 5.0 M Urea formulated in deionized water for 15 minutes. Gently rinse the CryoLoop in deionized water. Repeat three times. Allow the CryoLoop to air dry.

将 CryoLoop 浸泡在 0.5% 去离子水中的清洁剂溶液中 15 分钟。 在去离子水中轻轻冲洗 CryoLoop。 让 CryoLoop 风干。

如果需要更严格的清洁或不想冒残留洗涤剂残留的风险,请将 CryoLoop 浸泡在 5.0 M 去离子水中配制的尿素中 15 分钟。 在去离子水中轻轻冲洗 CryoLoop。 重复三遍。 让 CryoLoop 风干。

The potential benefits of cryogenic data collection 低温数据收集的潜在好处

The potential benefits of cryogenic data collection 低温数据收集的潜在好处

Reduced radiation damage.

Decreased thermal motion and disorder.

Potential for improved resolution.

Increased crystal lifetime.

Crystals can be stored and shipped.

减少辐射伤害。

减少热运动和紊乱。

提高分辨率的潜力。

增加晶体寿命。

晶体可以储存和运输。

Mounting thin crystals 安装晶体环方法

Mounting thin crystals 安装晶体环方法

To mount very thin crystals onto cryoloops, first dip the nylon loop into 0.5% Formvar solution (Fluka # 09819) to form a thin film. The film provides extra support for fragile crystals, and can result in much sharper reflections with just slightly higher background. To clean the loop, dip it in alcohol to dissolve the support. Two notes: (1) the technique works only for crystals grown without organic solvents, and (2) take precautions not to breathe vapor from the formvar solution–the solvent is 1,2 dichloroethane. Formvar is a standard support for electron microscopy grids.

要将非常薄的晶体安装到冷冻环上,首先将尼龙环浸入 0.5% Formvar 溶液 (Fluka # 09819) 中以形成薄膜。 该薄膜为易碎的晶体提供了额外的支撑,并且可以在背景稍高的情况下产生更清晰的反射。 要清洁环,请将其浸入酒精中以溶解支撑物。 两个注意事项:(1) 该技术仅适用于在没有有机溶剂的情况下生长的晶体,以及 (2) 采取预防措施,不要从 formvar 溶液中吸入蒸汽——溶剂是 1,2 二氯乙烷。 Formvar 是电子显微镜网格的标准支持。

PEGs for cryo 用于冷冻的 PEG(聚乙二醇)

PEGs for cryo 用于冷冻的 PEG(聚乙二醇)

High molecular PEGS are also good cryoprotectants. If crystals are obtained from relatively high concentration of PEGS (e.g., 30% of PEG 3350), you can cryoprotect them simply by raising the concentration of the PEG a little bit (e.g., 40% of PEG 3350).

高分子PEGS也是很好的冷冻保护剂。 如果晶体是从相对高浓度的 PEGS(例如 30% 的 PEG 3350)中获得的,您可以通过稍微提高 PEG 的浓度(例如 40% 的 PEG 3350)来冷冻保护它们。

参考文献

J. Appl. Cryst. (2006). 39, 244-251
Effects of cryoprotectant concentration and cooling rate on vitrification of aqueous solutions. V. Berejnov, N. S. Husseini, O. A. Alsaied and R. E. Thorne
Synopsis: Critical concentrations required for vitrification of aqueous solutions are determined for fourteen common cryoprotectants, for sample volumes ranging over four orders of magnitude and covering the range of interest in protein crystallography.

 

Trouble with protein storage 蛋白质储存问题

Trouble with protein storage 蛋白质储存问题

Try 3 to 5% 1,2-propanediol in the protein buffer as a substitute for glycerol to stabilize proteins. Annie Hassell, Glaxo Smithkline 2009

尝试在蛋白质缓冲液中加入 3% 到 5% 的 1,2-丙二醇作为甘油的替代品来稳定蛋白质。 安妮哈塞尔,葛兰素史克 2009