EMD-2173
Negative stain electron microscopy of a CSN-SCF~Nedd8/Skp2/Cks1 complex
EMD-2173
Single-particle25.0 Å
Deposition: 11/08/2012Map released: 03/10/2012
Last modified: 03/10/2012
Concentration: 0.2
mg/mL
Buffer
pH: 7.8
Details: 15 mM HEPES pH 7.8, 150 mM NaCl, 1% glycerol and 1 mM DTT
Details: 15 mM HEPES pH 7.8, 150 mM NaCl, 1% glycerol and 1 mM DTT
Staining
Type:
NEGATIVE
Details: Quantifoil grids (R2/2 Cu 400 mesh) coated with thin carbon floated from mica were glow-discharged for 30 seconds at 50 mA and 0.2 mbar vacuum. 3 ul purified samples were applied for 1 min to the grids. Following two brief buffer washes, the grids were stained with 2% uranyl acetate, gently blotted using filter paper and air-dried.
Details: Quantifoil grids (R2/2 Cu 400 mesh) coated with thin carbon floated from mica were glow-discharged for 30 seconds at 50 mA and 0.2 mbar vacuum. 3 ul purified samples were applied for 1 min to the grids. Following two brief buffer washes, the grids were stained with 2% uranyl acetate, gently blotted using filter paper and air-dried.
Microscope: FEI TECNAI F20
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 200 kV
Nominal magnification: 50000.0
Specimen holder model: SIDE ENTRY, EUCENTRIC
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 200 kV
Nominal magnification: 50000.0
Specimen holder model: SIDE ENTRY, EUCENTRIC
Image Recording
[1]
Detector category:
CCD
Detector model: GENERIC TVIPS (4k x 4k)
Average electron dose per image: 100 e/Å2
Details: Data was collected on a CCD camera
Detector model: GENERIC TVIPS (4k x 4k)
Average electron dose per image: 100 e/Å2
Details: Data was collected on a CCD camera
Details: See publication
Final
reconstruction
Resolution: 25.0
Å
(
BY AUTHOR)
Resolution method: FSC 0.5 CUT-OFF
Number of images used: 8055
Algorithm: OTHER
Details:
Resolution method: FSC 0.5 CUT-OFF
Number of images used: 8055
Algorithm: OTHER
Details:
⌯ Applied Symmetry
Point group:
C1
Software
[1]
| Name | Version | Details |
|---|---|---|
| IMAGIC, Spider, EMAN, in, house | - | - |
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: -
Details: ::::EMDATABANK.org::::EMD-2173::::
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: -
Details: ::::EMDATABANK.org::::EMD-2173::::
⬡ Geometry
| X | Y | Z | |
|---|---|---|---|
| Dimensions | 60 | 60 | 60 |
| Origin | -30 | -29 | -30 |
| Spacing | 60 | 60 | 60 |
| Voxel size | 3.47 Å | 3.47 Å | 3.47 Å |
Contour list
| Primary | Level | Source |
|---|---|---|
| True | 3.0 | AUTHOR |