| Method |
Hanging-drop vapor diffusion (manual) |
Hanging-drop vapor diffusion (manual) |
| Plate type |
EasyXtal 15-Well Tool + EasyXtal DropGuard Supports (Molecular Dimensions) |
EasyXtal 15-Well Tool + EasyXtal DropGuard Supports (Molecular Dimensions) |
| Temperature (K) |
277 |
293 |
| Protein concentration (mg ml−1/µM) |
9–11/228–280 |
3–5/82–137 |
| Protein buffer |
50 mM Tris pH 8.0, 250 mM NaCl, 5%(v/v) glycerol, 1 mM DTT |
50 mM Tris pH 8.0, 250 mM NaCl, 5%(v/v) glycerol, 1 mM DTT |
| Compound concentration |
Threefold molar excess of protein |
Threefold molar excess of protein |
| Protein–ligand incubation time |
1 h at 4°C |
1 h at 4°C |
| Reservoir solution |
100 mM imidazole pH 7.0–7.4, 25–30%(v/v) MPD |
75 mM imidazole pH 7.0–7.4, 8–14%(v/v) 2-propanol |
| Volume and ratio of drop |
1 µl + 1 µl |
1 µl + 1 µl |
| Volume of reservoir (µl) |
500 |
500 |
