EMD-2248 Experiments and Validation

negative stain single-particle reconstruction of conformation I of the Ltn1 E3 ubiquitin Ligase

Single particle reconstruction
Overview of EMD-2248

Map parameters

Minimum density: -1.252
Maximum density: 2.468
Average density: -0.008
Standard deviation: 0.146
Recommended contour level: 0.80 (author)

Sample information

Sample name: snapshot I of Ltn1 conformer
Protein: Ltn1

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Spider, Appion
Number of particles: 717
Number of class averages: 1
Reconstruction protocol: random-conical tilt reconstruction
CTF correction: each micrograph

Imaging

Session
Microscope model: FEI TECNAI F20
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 15.0
Nominal CS (mm): 2.00
Nominal magnification: 50000.0
Defocus max (nm): 2,600.00
Defocus min (nm): 1,200.00
Tilt max (degrees): 0.00
Tilt min (degrees): -55.00
Holder model: SIDE ENTRY, EUCENTRIC
Acquisition
Number of images: 489

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 0.01
Buffer: 190 mM NaCl, 20 mM Tris, 1mM BME
Staining procedure: 3 microliters of sample at a concentration of 0.01 mg/mL was applied to a C-flat grid with 2-micron-diameter holes overlaid by thin 1.5 nm continuous carbon. The specimen was stained with 2% uranyl formate 3 times, then let air-dry.
Specimen support: c-flat grids overlaid with thin (~1.5 nm) carbon
Vitrification
Apparatus: NONE
Cryogen: NONE