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CryoPro
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CryoPro
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Cryoprotectant reagent kit
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36 unique cryoprotectants
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Polyols, organics, oils, polymers, sugars, & salts
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Includes cryoprotectant tutorial
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Preformulated, ready to use
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As in selecting reagents for crystallization, the selection of a suitable cryoprotectant involves some trial and error as well as a screening.1-15 A suitable cryoprotectant, when mixed with the crystal and crystallization reagent, will cool to cryogenic temperature without ice formation and damage to the crystal. To assay for the proper concentration of cryoprotectant in the reagent used to grow the crystal, one can mix the cryoprotectant with the crystallization reagent and employ the desired cooling method (for example, place the solution in a CryoLoop™ and place the CryoLoop in a cryostream). Observe for ice formation either visually or with x-ray diffraction. Upon cooling, a transparent drop and x-ray diffraction pattern, free of powder diffraction rings or “ice rings” indicates success. The appearance of a cloudy drop or “ice rings” indicates an inappropriate cryoprotectant concentration or cryoprotectant. Incrementally increase the concentration and/or composition of the cryoprotectant serially 5 to 10% and repeat until the cooled drop remains clear while in the cryostream. Once a clear drop is achieved in the cryostream, this is typically a good starting point for cryopreservation of the crystal.
Some crystals can simply be dipped or washed quickly in a simple cryoprotectant such as 30% glycerol for successful cryopreservation. However, when all else fails, a rational assay of each cryoprotectant with incremental increases in cryoprotectant concentration as well as a test of mixtures (for example a mixture of sugars, or a sugar mixed with ethylene glycol) may be required to determine the best cryoprotectant for a crystal.
Each CryoPro - Cryoprotectant kit contains 36 unique cryoprotectants including organics, oils, polyols, salts, sugars, and polymers. 35 of the 36 reagents are supplied in 10 ml volumes, a single reagent L-(+)-2,3-butanol is supplied in a 0.2 ml aliquot. CryoPro is convenient and cost-effective.
Ready to use cryoprotectants are formulated using high purity reagents with ultra pure water and are sterile filtered. Many of the individual cryoprotectants are available as a 100 ml or 200 ml Optimize™ reagent. Please refer to the CryoPro Formulations pdf file to find the catalog number for the equivalent Optimize. Crystallization accessories are sold separately.
Formulations
Tube Number Cryoprotectant
1. 100% Glycerol
2. 100% Ethylene Glycol
3. 100% Polyethylene Glycol 200
4. 100% Polyethylene Glycol 400
5. 80% v/v Polyethylene Glycol 600
6. 60% w/v Polyethylene Glycol 4000
7. 50% w/v Polyvinylpyrrolidone K15
8. 100% (±)-2-Methyl-2,4-pentanediol
9. 6.0 M 1,6-Hexanediol
10. 100% Propylene Glycol
11. 100% Paratone-N
12. 100% Paraffin Oil
13. 100% NVH Oil
14. 100% Dimethyl Sulfoxide
15. 100% 2-Propanol
16. 100% Ethanol
17. 100% Methanol
18. 70% w/v D-(+)-Sucrose
19. 35% w/v meso-Erythritol
20. 70% w/v Xylitol
21. 15% w/v Inositol
22. 20% w/v D-(+)-Raffinose
23. 50% w/v D-(+)-Trehalose
24. 70% w/v D-(+)-Glucose
25. 100% 2,3-Butanediol
26. 100% L-(+)-2,3-Butanediol
27. 5.0 M Lithium Acetate
28. 10.0 M Lithium Chloride
29. 4.0 M Lithium Formate
30. 8.0 M Lithium Nitrate
31. 2.0 M Lithium Sulfate
32. 3.4 M Sodium Malonate pH 7.0
33. 3.5 M Magnesium Acetate
34. 5.0 M Sodium Chloride
35. 7.0 M Sodium Formate
36. 7.0 M Sodium Nitrate
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CAT NO |
NAME |
DESCRIPTION |
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| HR2-132 |
CryoPro |
10 ml, tube format |
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References
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| 1. | Boutron, P. (1987). Non-equilibrium formation of ice in aqueous solutions: efficiency of polyalcohol solutions for vitrification. In: Pegg, D.E. & Karow, A.M. Jr. (eds). The biophysics of organ prese |
| 2. | Garman, E.F., & Mitchell, E.P. (1996). Glycerol concentrations required for cryoprotection of 50 typical protein crystallization conditions. J. Appl. Cryst. 29, 584-587. |
| 3. | Garman, E.F., & Schneider, T.R. (1997). Macromolecular Cryocrystallography. J. Appl. Cryst. 30, 211-237. |
| 4. | Hope, H. (1988). Cryocrystallography of biological macromolecules: a generally applicable method. Acta Cryst. B 44, 22-26. |
| 5. | Kottke, T., & Stalke, D. (1993). Crystal handling at low temperatures. J. Appl. Cryst. 26, 615-619. |
| 6. | Kwong, P.D. Liu, Y. (1999). Use of cryoprotectants in combination with immiscible oils for flash cooling macromolecular crystals. J. Appl. Cryst. 32, 102-105. |
| 7. | Mehl, P. (1989). Experimental dissection of devitrification in aqueous solutions in 1,3-butanediol. Cryobiology. 26, 567-568. |
| 8. | Parkin, S., & Hope, H. (1998). Macromolecular cryocrystallography: Cooling, mounting, storage and transportation of crystals. J. Appl. Cryst. 31, 945-953. |
| 9. | Petcock, J.M., Wang, Y.-F., DuBois, G.C., Harrison, R.W., & Weber, I.T. (2001). Effects of different post-crystallization soaking conditions on the diffraction of Mtcp1 crystals. Acta Cryst. D57, 763- |
| 10. | Petsko, G.A. (1975). J. Mol. Biol. 96, 381-392. |
| 11. | Rodgers, D.W. (1994). Cryocrystallography. Structure. 2, 1135-1140. 12. Schneider, T.R. (1997). Cryocrystallography of biological macromolecules. Acta Physica Polonica A. 91, 739-744. |
| 12. | Schneider, T.R. (1997), Cryocrystallography of biological macromolecules. Acta Physica Polonica A. 91, 739-744. |
| 13. | Teng, T.-Y. (1990). J. Appl. Cryst. 23, 387-391. |
| 14. | Walker, L.J., Moreno, P.O., Hope, H. (1998). Cryocrystallography: effect of cooling medium on sample cooling rate. J. Appl. Cryst. 31, 954-965. |
| 15. | Watenpaugh, K.D. (1991). Curr. Op. Struct. Biol. 1, 1012. |
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