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Title:Single particle analysis of recombinant human anaphase promoting complex (APC/C)
Authors:Zhang Z, Yang J, Kong EH, Chao WCH, Morris EP, da Fonseca PCA, Barford D
Sample:recombinant human APC/C
Method:Single particle reconstruction (20 angstroms resolution)
Red flagLatest update:2012-12-26
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Sample
Sample name: recombinant human APC/C
Theoretical molecular weight of the sample: 1.2
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinApc1monomertrueHomo sapiens
2proteinApc5monomertrueHomo sapiens
3proteinApc15monomertrueHomo sapiens
4proteinApc10monomertrueHomo sapiens
5proteinApc8monomertrueHomo sapiens
6proteinApc13monomertrueHomo sapiens
7proteinApc2monomertrueHomo sapiens
8proteinApc4monomertrueHomo sapiens
9proteinApc11monomertrueHomo sapiens
10proteinApc3monomertrueHomo sapiens
11proteinApc16monomertrueHomo sapiens
12proteinCdc26monomertrueHomo sapiens
13proteinApc7monomertrueHomo sapiens
14proteinApc6monomertrueHomo sapiens
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.0 mg/mL20 mM HEPES NaOH pH 8.0, 200 mM NaCl, 2 mM DTTgrids stained with 2% (w/v) uranyl acetateQuantifoil R1.2/1.3 grids coated with a thin layer of carbon
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 FIELD mm1200 nm nm50000FIELD EMISSION GUNTietz F415 CCD mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC°° eV K K K mrad100 e/Å201-JAN-2012
Processing
Software:Spider, Imagic
Number of particles:5639
Resolution by author:20 Å
Resolution method:FSC
Processing details:Particles were manually selected using the EMAN boxer software. The images were analysed using a combination of IMAGIC and Spider software.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
μm/pixelon recording each adjacent boxes of 2x2 pixels were averaged
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails