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Title:map of the complex of the HA7 Fab with avian flu virus hemagglutinin by negative staining EM
Authors:Du L, Jin L, Zhao G, Sun S, Li J, Li Y, Zheng B, Liddington CL, Zhou Y, Jiang S
Sample:the complex of Fab Fragment of HA7 monoclonal antibody and avian influenza virus hemagglutinin
Method:Single particle reconstruction (18.0 angstroms resolution)
Red flagLatest update:2013-07-24
Sample
Sample name: the complex of Fab Fragment of HA7 monoclonal antibody and avian influenza virus hemagglutinin
Oligomeric state: one trimeric hemagglutinin binds to three Fab
Experimental molecular weight of the sample: 0.37
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinFab Fragment of HA7 monoclonal antibodytrimertrueMus musculus
2proteinAvian influenza virus hemagglutinintrimertrueunidentified influenza virusIPR001364
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.4 mg/mL50mM NaCl, 10mM Tris-HCL,1mM CaCl2, pH7.4Grids with adsorbed protein floated on 2% w/v uranyl formate for 30 seconds300 mesh copper grid with carbon support, glow discharged in vacuumed air
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1200 nm3600 nm70000FIELD EMISSION GUNGENERIC TVIPS (4k x 4k) mmObjective lens astigmatism was corrected at 70,000 times magnification.

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC eV K K K mrad50 e/Å207-AUG-2011
Processing
Protocol:Cross-common lines
Software:EMAN2
CTF correction:Each particle
Number of particles:3172
Resolution by author:18.0 Å
Resolution method:FSC 0.5
Processing details:C3 symmetry was imposed in refinement. Image processing was done using EMAN2.The particles were selected by a semi-automatic selection using e2boxer.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
53 μm/pixel8
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails