Using CBTP one can use a single buffer system to screen pH 2.5 to 9.5 by varying the ratio of Citric acid to BIS-TRIS propane. Although the two buffer reagents are made available separately for convenient, individual replacement, they are designed to be used together, with the supplied CBTP buffer titration table to create a crystallization buffer system covering pH 2.5 to 9.5.

HR2-831 Measured pH range: 1.2 – 1.5 at 25°C
HR2-831 Measured Conductivity Range: 7.7 – 8.7 mS/cm at 25°C

HR2-833 Measured pH range: 11.2 – 11.5 at 25°C
HR2-833 Measured Conductivity Range: 298 – 317 µS/cm at 25°C
HR2-833 Measured Refractive Index Range: 1.38000 – 1.38076 at 20°C

1.0 M Citric acid

1.0 M BIS-TRIS propane

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: O-(2-Aminopropyl)-O′-(2-methoxyethyl)polypropylene glycol 500 and Polypropylene glycol 500 mono-2-aminoethyl mono-2-methoxyethyl ether
CH₃OCH₂CH₂O[CH(CH₃)CH₂O]_nCH₂CH(NH₂)CH₃
M_r ~600 
CAS Number [77110-54-4] 
MDL Number MFCD00147774 
PubChem Substance ID 24846306 
Clear / Colorless / Light Yellow / Light Brown 
Jeffamine^® M-600^® is utilized in Additive Screen, Additive Screen HT, Crystal Screen 2, Crystal Screen HT, Index, Index HT, PEGRx 1 and PEGRx HT crystallization screens 
HR2-501 is titrated to pH 7.0 using HCl and is sterile filtered 

HR2-501 is titrated to pH 7.0 using HCl
HR2-501 Measured Conductivity Range: 8.8 – 10.3 mS/cm at 25°C 
HR2-501 Measured Refractive Index Range: 1.40707 – 1.40988 at 20°C 

HR2-503 is not titrated and the measured pH is >12.0 
HR2-503 Measured Conductivity Range: 0.0 – 0.1 µS/cm at 25°C 
HR2-503 Measured Refractive Index Range: 1.44499 – 1.44565 at 20°C 

Hampton Research recommends using HR2-501 to reproduce and optimize the following Hampton Research screen reagents: 
-Crystal Screen 2 reagent 18 
0.01 M Iron(III) chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 10% v/v Jeffamine^® M-600^® 

-Index reagent 38 
30% v/v Jeffamine^® M-600^® pH 7.0, 0.1 M HEPES pH 7.0 

Jeffamine is a registered trademark of the Huntsman Petrochemical Corporation. 
M-600 is a registered trademark of Texaco.

50% v/v Jeffamine M-600 pH 7.0

100% Jeffamine M-600 (untitrated pH >12)

1. Crystallization and X-ray diffraction analysis of an all-RNA U39C mutant of the minimal hairpin ribozyme. V. Grum-Tokars, M. Milovanovic and J. E. Wedekind. Acta Cryst. (2003). D59, 142-145.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: 2-[4-(2-Hydroxyethyl)-1-piperazinyl]ethanesulfonic acid or 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid or N-(2-Hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)
C₈H₁₈N₂O₄S
M_r 238.31
CAS Number [7365-45-9]
EC Number 230-907-9
Beilstein Registry Number 883043
Merck 14,4654
RTECS TL6809000
MDL Number MFCD00006158
PubChem Substance ID 24895616
Purity ≥ 99.5%
pKa 7.5 at 25°C

Useful pH range of HEPES buffer 6.8 – 8.2

Titrate HR2-585 to the desired pH between 6.8 – 8.2 using NaOH
HR2-585 Measured pH Range: 5.4 – 5.5 at 25°C
HR2-585 Measured Conductivity Range: 160.1 – 236.9 μS/cm at 25°C
HR2-585 Measured Refractive Index Range: 1.37093 – 1.37131 at 20°C

HR2-785 is titrated to pH 7.0 using NaOH
HR2-785 Measured Conductivity Range: 5.8 – 7.0 mS/cm at 25°C
HR2-785 Measured Refractive Index Range: 1.37133 – 1.37176 at 20°C

HR2-729 is titrated to pH 7.5 using NaOH
HR2-729 Measured Conductivity Range: 11.6 – 13.6 mS/cm at 25°C
HR2-729 Measured Refractive Index Range: 1.37177 – 1.37290 at 20°C

Insoluble Matter 0.01% max
DNase None Detected
RNase None Detected
Protease None Detected
Heavy Metals 1 ppm max
As 5 ppm max
Cu 5 ppm max
Fe 5 ppm max
Pb 5 ppm max
SO₄ 0.005% max

1.0 M HEPES

1.0 M HEPES pH 7.0

1.0 M HEPES pH 7.5

1. Crystallization and preliminary X-ray crystallographic studies of native elastase from North Atlantic salmon (Salmo salar). Berglund, GI; Snalas, AO; Hansen, LK. Acta Crystallogr D 51, 393- 394, 1995.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: None
C[CH₂(OCH₂CH₂)_nOH]₄
Average M_n ~ 270
CAS Number [30599-15-6]
EC Number 500-071-2
Density 1.2 g/mL at 25°C (literature)
Boiling point >300°C (literature)
Refractive index n20/D 1.481

Measured Conductivity Range: 36.1 – 61.3 µS/cm at 25°C
Measured Refractive Index Range: 1.39627 – 1.39719 at 20°C

1. Acta Cryst. (2002). D58, 306-309. Pentaerythritol propoxylate: a new crystallization agent and cryoprotectant induces crystal growth of 2-methylcitrate dehydratase. A. M. Gulick, A. R. Horswill, J. B. Thoden, J. C. Escalante-Semerena and I. Rayment.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: HEPES sodium salt, or 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid sodium salt, or N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) sodium salt
C₈H₁₇N₂NaO₄S
M_r 260.29
CAS [75277-39-3]
EC Number 278-169-7
MDL Number MFCD00036463
PubChem Substance ID 24895780
pKa 7.5 at 25°C
Useful pH Range of HEPES sodium: 6.6 – 8.2

HR2-733 is titrated to pH 7.5 using HCl
HR2-733 measured Conductivity range: 41.7 – 43.4 mS/cm at 25°C
HR2-733 measured Refractive Index range: 1.37577 – 1.37766 at 20°C

HR2-577 is the free base of HEPES sodium and is to be titrated to the desired pH using HCl
HR2-577 measured pH Range: 10.2 – 12.1 at 25°C
HR2-577 measured Conductivity range: 25.9 – 27.2 mS/cm at 25°C
HR2-577 measured Refractive Index range: 1.37097 – 1.37298 at 20°C

HR2-931-01 – 1.0 M HEPES sodium pH 6.8 is a Custom Shop reagent and is used to reproduce and optimize Crystallization Reagents for use with the Silver Bullets kit.

1.0 M HEPES sodium

1.0 M HEPES sodium pH 7.5

1. Crystallization and preliminary X-ray characterization of GM2-activator protein. C. S. Wright and S.-C. Li. Acta Cryst. (1997). D53, 211-212.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: Pentaerythritol ethoxylate
C[CH₂(OCH₂CH₂)_nOH]₄, N = 3.75
Average M_n ~ 797
CAS Number [30599-15-6]
EC Number 500-071-2
MDL Number MFCD00197884
Boiling point >300°C (literature)

Measured Conductivity Range: 3.2 – 11.4 µS/cm at 25°C
Measured Refractive Index Range: 1.40400 – 1.41578 at 20°C

Pentaerythritol ethoxylate (15/4 EO/OH) consists of a mixture of different compounds. The “15” indicates that there are 15 different chain length compounds in the mixture with an average chain length of 4.

50% v/v Pentaerythritol ethoxylate (15/4 EO/OH)

1. Acta Cryst. (2002). D58, 306-309. Pentaerythritol propoxylate: a new crystallization agent and cryoprotectant induces crystal growth of 2-methylcitrate dehydratase. A. M. Gulick, A. R. Horswill, J. B. Thoden, J. C. Escalante-Semerena and I. Rayment.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: 1,3-Diaza-2,4-cyclopentadiene or Glyoxaline
C₃H₄N₂
M_r 68.08
CAS Number [288-32-4]
EC Number 206-019-2
Beilstein Registry Number 103853
Merck 14,4912
RTECS NI3325000
MDL Number MFCD00005183
PubChem Substance ID 24880372
Purity ≥ 99.5%
UN Number 2923 8/PG 3

pKa = 6.95
The useful pH range of an Imidazole can be 6.2 – 7.8

HR2-819 is titrated to pH 7.0 using HCl
HR2-819 Measured Conductivity Range: 45.4 – 50.4 mS/cm at 25°C
HR2-819 Measured Refractive Index Range: 1.34836 – 1.34904 at 20°C

Titrate HR2-573 to the desired pH using HCl
HR2-573 Measured pH Range: 10.5 – 10.7 at 25°C
HR2-573 Measured Conductivity Range: 71.1 – 101.5 µS/cm at 25°C
HR2-573 Measured Refractive Index Range: 1.34420 – 1.34440 at 20°C

Ca  ≤0.001%
Cd  ≤0.0005%
Cl   ≤0.005%
Co  ≤0.0005%
Cr  ≤0.0005%
Cu  ≤0.0005%
Fe  ≤0.0005%
K   ≤0.005%
Mg ≤0.0005%
Na ≤0.005%
Ni  ≤0.0005%
Pb  ≤0.0005%
SO₄ ≤0.005%
Zn  ≤0.0005%

1.0 M Imidazole

1.0 M Imidazole pH 7.0

1. Structure of apo acyl carrier protein and a proposal to engineer protein crystallization through metal ions. X. Qiu and C. A. Janson. Acta Cryst. (2004). D60, 1545-1554 [doi:10.1107/S0907444904015422].

2. Protein crystallization by surface entropy reduction: optimization of the SER strategy. D. R. Cooper, T. Boczek, K. Grelewska, M. Pinkowska, M. Sikorska, M. Zawadzki and Z. Derewenda. Acta Cryst. (2007). D63, 636-645 [doi:10.1107/S0907444907010931].

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: None
C[CH₂[OCH₂CH(CH₃)]_nOH]₄ N~ 5
M_r ~426
CAS Number [9051-49-4]
EC Number 500-030-9
MDL Number MFCD00197885
PubChem Substance ID 24866238
Density of 100% solution 1.05 g/ml at 25°C
Refractive index of 100% solution n20/D 1.464
Boiling point of 100% solution >300°C (literature)

Measured Conductivity Range: 4.2 – 5.4 µS/cm at 25°C
Measured Refractive Index Range: 1.40400 – 1.40447 at 20°C

50% v/v Pentaerythritol propoxylate (5/4 PO/OH)

1. Acta Cryst. (2002). D58, 306-309. Pentaerythritol propoxylate: a new crystallization agent and cryoprotectant induces crystal growth of 2-methylcitrate dehydratase. A. M. Gulick, A. R. Horswill, J. B. Thoden, J. C. Escalante-Semerena and I. Rayment.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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Synonyms: MES or 2-(N-morpholino)ethanesulfonic acid or 4-morpholineethanesulfonic acid monohydrate
C₆H₁₃NO₄S • H₂O
M_r 213.25
CAS Number [145224-94-8]
EC Number 224-632-3
Beilstein Registry Number 329824236
RTECS QE3479500
MDL Number MFCD00149409
PubChem Substance ID 24885996
Purity ≥ 99.0%
pKa 6.1 at 25°C

Solutions are stable at 2-8°C for months. Autoclaving is not recommended for any sulfonic acid buffer. When MES solutions are autoclaved, they turn yellow (although pH does not change measurably). The identity of the yellow breakdown product is unknown. When MES solutions are stored at room temperature, they can turn yellow within six months.

Useful pH range of MES 5.2 – 7.1

Titrate HR2-587 to the desired pH using NaOH
HR2-587 Measured pH Range: 3.2 – 3.4 at 25°C
HR2-587 Measured Conductivity Range: 166.4 – 2440 µS/cm at 25°C
HR2-587 Measured Refractive Index Range: 1.34655 – 1.34731 at 20°C

HR2-787 has been titrated to pH 6.5 using NaOH
HR2-787 Measured Conductivity Range: 19.2 – 25.6 mS/cm at 25°C
HR2-787 Measured Refractive Index Range: 1.36113 – 1.36251 at 20°C

HR2-943-07 – MES monohydrate pH 5.8 – 1.0 M solution is a Custom Shop reagent and is used to reproduce and optimize Crystallization Reagents for use with the Silver Bullets kit.

Impurities:
Endotoxin ≤ 50 EU/g
Yeast and Mold ≤ 100 CFU/g
Bioburden ≤ 100 CFU/g
Al ≤ 5.0 ppm
As ≤ 3.0 ppm
Ba ≤ 5.0 ppm
Bi ≤ 5.0 ppm
Ca ≤ 20.0 ppm
Cd ≤ 5.0 ppm
Co ≤ 5.0 ppm
Cr ≤ 5.0 ppm
Cu ≤ 5.0 ppm
Fe ≤ 5.0 ppm
Li ≤ 5.0 ppm
Mg ≤ 5.0 ppm
Mn ≤ 5.0 ppm
Mo ≤ 5.0 ppm
Na ≤ 20.0 ppm
Ni ≤ 5.0 ppm
Pb ≤ 5.0 ppm
Sb ≤ 5.0 ppm
Zn ≤ 5.0 ppm

0.5 M MES monohydrate

1.0 M MES monohydrate pH 6.5

1. Crystal structures of mutant Pseudomonas aeruginosa p-hydroxybenzoate hydroxylases: the Tyr201Phe, Tyr385Phe, and Asn300Asp variants. Lah, MS; Palfey, BA; Schreuder, HA; Ludwig, ML. Biochemistry 33, 1555- 1564, 1994.

Hampton Research, first in crystallization since 1991, developing and delivering crystallization and optimization screens, reagents, plates, and other tools for the crystallization of biological macromolecules, including proteins (antibody), peptides (insulin), and nucleic acids (DNA).

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