Jun 20, 2019
Buffer Recipes for Protein Purification
- 1University of California San Francisco
Protocol Citation: Annie Kwon 2019. Buffer Recipes for Protein Purification. protocols.io https://dx.doi.org/10.17504/protocols.io.4e8gthw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 20, 2019
Last Modified: June 20, 2019
Protocol Integer ID: 24768
Keywords: Buffers, protein purification
Equilibration buffer
Equilibration buffer
25 mM HEPES, 100 mM NaCl, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 1.461 g | 2.922 g | 5.844 g | 11.688 g | 23.376 g |
Dissolve in water; bring pH up to 7.5; store at 4°C
25 mM HEPES, 500 mM NaCl, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 7.305 g | 14.61 g | 29.22 g | 58.44 g | 116.88 g |
Dissolve in water; bring pH up to 7.5; store at 4°C
Lysis buffer
Lysis buffer
50 mM Na/K phosphate buffer, 500 mM NaCl, 2.5% glycerol, pH 8.0
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| NaH2PO4 • H2O (MW=137.9923) | 0.0690 g | 0.1380 g | 0.2760 g | 0.5520 g | 1.1039 g | |
| K2HPO4 (MW=174.18) | 2.0902 g | 4.1803 g | 8.3607 g | 16.7213 g | 33.4426 g | |
| NaCl (MW=58.44) | 7.305 g | 14.61 g | 29.22 g | 58.44 g | 116.88 g | |
| glycerol | 6.25 mL | 12.5 mL | 25 mL | 50 mL | 100 mL |
Dissolve in water; bring pH up to 8.0; store at 4°C
Wash buffer
Wash buffer
25 mM HEPES, 1M NaCl, 10 mM imidazole, 10% glycerol, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 14.61 g | 29.22 g | 58.44 g | 116.88 g | 233.76 g | |
| imidazole (MW=68.08) | 0.1702 g | 0.3404 g | 0.6808 g | 1.3616 g | 2.7232 g | |
| glycerol | 25 mL | 50 mL | 100 mL | 200 mL | 400 mL |
Dissolve in water; bring pH up to 7.5; store at 4°C
25 mM HEPES, 1M NaCl, 10 mM imidazole, 10% glycerol, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 14.61 g | 29.22 g | 58.44 g | 116.88 g | 233.76 g | |
| imidazole (MW=68.08) | 0.3404 g | 0.6808 g | 1.3616 g | 2.7232 g | 5.4464 g | |
| glycerol | 25 mL | 50 mL | 100 mL | 200 mL | 400 mL |
Dissolve in water; bring pH up to 7.5; store at 4°C
Elution buffer
Elution buffer
25 mM HEPES, 100 mM NaCl, 300 mM imidazole, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 1.461 g | 2.922 g | 5.844 g | 11.688 g | 23.376 g | |
| imidazole (MW=68.08) | 5.106 g | 10.212 g | 20.424 g | 40.848 g | 81.696 g |
Dissolve in water; bring pH up to 7.5; store at 4°C
25 mM HEPES, 500 mM NaCl, 300 mM imidazole, pH 7.5
| 250 mL | 500 mL | 1 L | 2 L | 4 L | ||
| HEPES (MW=238.31) | 1.4894 g | 2.9789 g | 5.9578 g | 11.9155 g | 23.831 g | |
| NaCl (MW=58.44) | 7.305 g | 14.61 g | 29.22 g | 58.44 g | 116.88 g | |
| imidazole (MW=68.08) | 5.106 g | 10.212 g | 20.424 g | 40.848 g | 81.696 g |
Dissolve in water; bring pH up to 7.5; store at 4°C