The Protein Purification Facility
The Alexander Silberman Institute of Life Sciences
The Hebrew University of Jerusalem
Dr. Mario Lebendiker
mariol@mail.ls.huji.ac.il  Tel: 972-2-6586920  FAX: 972-2-6758963

Recommended Buffers for Anion Exchange Chromatography

Recommended buffer substances for anion exchange chromatography. According to Amersham Biosciences catalogue
pH interval  Substance  Conc. (mM)  Counter-ion  pKa (25 °C)  d(pKa)/dT (°C) 
4.5-5.0  N-Methylpiperazine*  20  Cl- 4.75  -0.015 
5.0-6.0  Piperazine*  20  Cl- or HCOO- 5.68  -0.015 
5.5-6.0  L-Histidine*  20  Cl- 6.15   
5.8-6.4  bis-Tris*  20  Cl- 6.50  -0.017 
6.4-7.3  bis-Tris propane*  20  Cl- 6.80   
7.3-7.7  Triethanolamine*  20  Cl- or CH3COO- 7.77  -0.020 
7.5-8.0  Tris*  20  Cl- 8.16  -0.028 
8.0-8.5  N-Methyldiethanolamine*  20  SO42- 8.54  -0.028 
8.0-8.5  N-Methyldiethanolamine*  50  Cl- or CH3COO- 8.54  -0.028 
8.4-8.8  Diethanolamine*  20 at pH 8.4  Cl- 8.88  -0.025 
    50 at pH 8.8       
8.5-9.0  1,3-Diaminopropane*  20  Cl- 8.64  -0.031 
9.0-9.5  Ethanolamine*  20  Cl- 9.50  -0.029 
9.5-9.8  Piperazine*  20  Cl- 9.73  -0.026 
9.8-10.3  1,3-Diaminopropane*  20  Cl- 10.47  -0.026 
10.6-11.6  Piperidine  20  Cl- 11.12  -0.031 
* Recommended on the basis of experiments in Amersham Biosciences laboratories. 

 
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Dr. Mario Lebendiker The Protein Purification Facility
The Alexander Silberman Institute of Life Sciences,    The Hebrew University of Jerusalem
mariol@mail.ls.huji.ac.il  Tel: 972-2-6586920  FAX: 972-2-6758963

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