| Fragment |
279–568 |
47–320 |
| Expression temperature (°C) |
22 |
18 |
| [IPTG] (mM) |
0.2 |
0.2 |
| Lysis buffer |
25 mM Tris–HCl pH 7.8, 300 mM NaCl, 5 mM MgCl2, 5 mM β-mercaptoethanol, 12% glycerol |
50 mM Tris–HCl pH 8.0, 500 mM NaCl, 10% glycerol, 50 mM arginine, 50 mM glutamic acid, 5 mM TCEP |
| Affinity column |
TALON Superflow |
Nickel–nitriloacetic acid resin |
| Activity buffer |
25 mM Tris–HCl pH 7.8, 300 mM NaCl, 5 mM MgCl2, 8% glycerol |
50 mM Tris–HCl pH 8.0, 500 mM NaCl, 10% glycerol |
| SEC buffer |
20 mM HEPES pH 7.5, 300 mM NaCl, 5 mM MgCl2, 12% glycerol, 5 mM β-mercaptoethanol |
50 mM Tris–HCl pH 8.0, 500 mM NaCl, 10% glycerol, 50 mM arginine, 50 mM glutamic acid, 5 mM TCEP |
