EMD-5898 Experiments and Validation

Cryo EM density of microtubules stabilized by GMPCPP, without kinesin

Single particle reconstruction
Overview of EMD-5898
Sample name: microtubule stabilized by GMPCPP, without kinesin
Organism: Sus scrofa
Related EM entries by publication: EMD-5895, EMD-5896, EMD-5897, EMD-5899

Map parameters

Recommended contour level: 3.20 (emdb)
Number of grid points: 110 × 66 × 162
Voxel size: 1.74 × 1.74 × 1.74 Å
Minimum density: -1.569
Maximum density: 4.923
Average density: 0.382
Standard deviation: 1.521

Sample information

Sample name: microtubule stabilized by GMPCPP, without kinesin
Proteins: Alpha tubulin, Beta tubulin

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: FREALIGN
Number of particles: 11394
Reconstruction protocol: projection matching
CTF correction: ctftilt

Imaging

Session
Microscope model: FEI TITAN
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 25.0
Nominal CS (mm): 2.70
Nominal magnification: 72000.0
Defocus max (nm): 3,500.00
Defocus min (nm): 1,400.00
Tilt max (degrees): 0.00
Tilt min (degrees): 0.00
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 43
Scanner model: NIKON SUPER COOLSCAN 8000
Sampling interval (μm): 6.3499999999999996

Specimen

Preparation
Specimen state: helicalArray
Specimen concentration (mg/mL): 0.25
Buffer: 80mM PIPES, 1mM EGTA, 1mM MgCl2, 1mM DTT, 0.05% Nonidet P-40
Specimen support: 400 mesh C-flat 1.2/1.3, glow discharged in Edwards carbon evaporator
Aggregation parameters (helical parameters)
Handedness: LEFT HANDED
Phi increment (degrees): 25.75
Azimuthal (Z) increment (Å): 8.9000000000000004
Rotational symmetry: C1
Vitrification
Apparatus: FEI VITROBOT MARK II
Cryogen: ETHANE
Humidity: 90
Protocol: The grid was blotted for 2 seconds before plunging.