| Method |
Vapor diffusion, sitting drop |
Vapor diffusion, sitting drop |
Vapor diffusion, sitting drop |
| Temperature (K) |
290 |
290 |
287 |
| Protein concentration (mg ml−1) |
25 |
25 |
25 |
| Protein buffer composition |
20 mM HEPES pH 7.0, 300 mM NaCl, 5%(v/v) glycerol, 1 mM TCEP |
20 mM HEPES pH 7.0, 300 mM NaCl, 5%(v/v) glycerol, 1 mM TCEP |
20 mM HEPES pH 7.0, 300 mM NaCl, 5%(v/v) glycerol, 1 mM TCEP |
| Composition of reservoir solution |
100 mM CHES–NaOH pH 9.5, 30%(w/v) PEG 3000 |
1 M LiCl, 100 mM sodium acetate, 30%(w/v) PEG 6000 |
20%(w/v) PEG 3350, 200 mM potassium nitrate |
| Volume and ratio of drop |
0.4 µl:0.4 µl |
0.4 µl:0.4 µl |
0.5 µl:0.5 µl |
| Volume of reservoir (µl) |
50 |
50 |
50 |
| Cryoprotectant |
Al’s oil |
30 s soak in cryo/phasing solution 1 [4.5 µl reservoir + 0.5 µl 2.5 M NaI, 20%(v/v) ethylene glycol], 30 s soak in cryo/phasing solution 2 [4 µl reservoir + 1 µl 2.5 M NaI, 20%(v/v) ethylene glycol] |
Soak in 10 mM threonine, 20%(w/v) PEG 3350, 200 mM potassium nitrate, 20% ethylene glycol |
