ELECTRON MICROSCOPY

Sample

Ternary complex of human A1AT-NH and Chaetomium thermophilum Endoplasmic reticulum degradation enhancing mannosidase (CtEDEM) plus Endoplasmic reticulum degradation enhancing protein disulfide isomerase (CtPDI)

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name PROPANE
Sample Vitrification Details 3 uL of protein were added to the grid for 1 minute, at 4 degrees C with 100% humidity, before blotting with Whatman No1 filter paper (Sigma-aldrich, Germany) and washing three times with 20 uL drops of ultrapure water. Blot force 10 for 3 s before plunge freezing immediately into liquid ethane. A 30 second wait time after sample application before plunge freezing.
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 0765
Reported Resolution (Å) 3.46
Resolution Method FSC 0.143 CUT-OFF
Other Details ?
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space ?
Refinement Protocol ?
Refinement Target ?
Overall B Value ?
Fitting Procedure ?
Details ?
Data Acquisition
Detector Type GATAN K3 (6k x 4k)
Electron Dose (electrons/Å2) 11.15
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1000
Maximum Defocus (nm) 2500
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS 2.7
Imaging Mode BRIGHT FIELD
Specimen Holder Model FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification ?
Calibrated Magnification 105000
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION Topaz ?
MODEL REFINEMENT PHENIX 1.21.2_5419
IMAGE ACQUISITION EPU ?
CLASSIFICATION cryoSPARC ?
RECONSTRUCTION cryoSPARC ?
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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