8. Ultrapure water.
9. 10–14 K MWCO dialysis tubing, 1 cm flat width.
10. Dialysis clips (3 cm in length or larger).
11. Incubators (heating/cooling).
12. Fumehood for work with phospholipids in chloroform.
2.2 Negative
Staining of 2D
Crystallization Trials
1. 400-mesh, carbon-coated copper grids.
2. 1–2% uranyl acetate solution.
3. Forceps (Dumont #5) and/or anticapillary forceps (Dumont
#N5AC).
4. Whatman #4 filter paper.
5. A glow-discharge unit.
6. Grid storage boxes.
7. Desiccator cabinet.
2.3 TEM Screening
of Crystallization Trials
1. 80–120 kV transmission electron microscope (TEM) equipped
with suitable detector for FFT assessment.
2.4 Cryo-EM Grid
Preparation, Screening
and Data Collection
1. 400-mesh copper grids or 400-mesh molybdenum grids.
2. Carbon-coated mica.
3. Anticapillary forceps.
4. 4% trehalose (alternatively, 2% buffered tannin solution or 2%
glucose).
5. Whatman #4 filter paper.
6. Liquid nitrogen.
7. Liquid nitrogen dewar.
8. Protective gloves and face shield or protective glasses for work
with liquid nitrogen.
9. MilliporeSigma DAWP 0.65 μm filter membrane filter paper
(for peel-blot).
10. Parafilm M laboratory film (for peel-blot).
11. 600-mesh copper grids or 600-mesh molybdenum grids (for
peel-blot).
12. 120 or 200 kV screening cryo-EM (once crystals are available
and ready for cryo-EM).
13. 300 kV cryo-EM for data collection.
2.5 Image
Processing
1. PC, Mac, or workstation.
2. Focus software package: https://focus.c-cina.unibas.ch/
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Matthew C. Johnson et al.
9. 10–14 K MWCO dialysis tubing, 1 cm flat width.
10. Dialysis clips (3 cm in length or larger).
11. Incubators (heating/cooling).
12. Fumehood for work with phospholipids in chloroform.
2.2 Negative
Staining of 2D
Crystallization Trials
1. 400-mesh, carbon-coated copper grids.
2. 1–2% uranyl acetate solution.
3. Forceps (Dumont #5) and/or anticapillary forceps (Dumont
#N5AC).
4. Whatman #4 filter paper.
5. A glow-discharge unit.
6. Grid storage boxes.
7. Desiccator cabinet.
2.3 TEM Screening
of Crystallization Trials
1. 80–120 kV transmission electron microscope (TEM) equipped
with suitable detector for FFT assessment.
2.4 Cryo-EM Grid
Preparation, Screening
and Data Collection
1. 400-mesh copper grids or 400-mesh molybdenum grids.
2. Carbon-coated mica.
3. Anticapillary forceps.
4. 4% trehalose (alternatively, 2% buffered tannin solution or 2%
glucose).
5. Whatman #4 filter paper.
6. Liquid nitrogen.
7. Liquid nitrogen dewar.
8. Protective gloves and face shield or protective glasses for work
with liquid nitrogen.
9. MilliporeSigma DAWP 0.65 μm filter membrane filter paper
(for peel-blot).
10. Parafilm M laboratory film (for peel-blot).
11. 600-mesh copper grids or 600-mesh molybdenum grids (for
peel-blot).
12. 120 or 200 kV screening cryo-EM (once crystals are available
and ready for cryo-EM).
13. 300 kV cryo-EM for data collection.
2.5 Image
Processing
1. PC, Mac, or workstation.
2. Focus software package: https://focus.c-cina.unibas.ch/
230
Matthew C. Johnson et al.