EMD-2241 Experiments and Validation

Negative stain reconstruction of PG9 Fab in complex with HIV-1 SOSIP gp140 trimer

Single particle reconstruction
Overview of EMD-2241
Sample name: Fab fragment of broadly neutralizing antibody PG9 bound to BG505 SOSIP gp140 HIV-1 trimer
Organisms: Homo sapiens, Human immunodeficiency virus 1

Map parameters

Minimum density: -1.92
Maximum density: 10.456
Average density: 0.00
Standard deviation: 0.538
Recommended contour level: 1.03 (author)

Sample information

Sample name: Fab fragment of broadly neutralizing antibody PG9 bound to BG505 SOSIP gp140 HIV-1 trimer
Proteins: Broadly Neutralizing Antibody PG9, HIV-1 SOSIP.664

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Xmipp, EMAN
Number of particles: 33431
CTF correction: Not corrected

Imaging

Session
Microscope model: FEI TECNAI F20
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 30.0
Nominal magnification: 100000.0
Defocus max (nm): 900.00
Defocus min (nm): 500.00
Tilt max (degrees): 55.00
Tilt min (degrees): 0.00
Holder model: SIDE ENTRY, EUCENTRIC
Details: Images taken from 0 to -55 degrees in 5 degree tilt increments.
Acquisition
Number of images: 1283
Sampling interval (μm): 0.109
Details: Images recorded on CCD

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 0.03
Buffer: 50 mM Tris-HCl, 150 mM NaCl
Staining procedure: 30 sec Nano-W
Specimen support: 400 Cu mesh grid with carbon support, glow discharged at 20 mA for 30 seconds
Vitrification
Apparatus: NONE
Cryogen: NONE