SAXS profiles for both Nucb2s in solution were obtained by the use of XEUSS 2.0 SAXS/WAXS system (Xenocs, Grenoble, France). The ggNucb2 and hsNucb2 samples (protein concentration of 3.5 mg/ml) with 5 mM EDTA or 10 mM Ca2+, 100 µM Zn2+ and 10 mM Mg2+ and reference sample (BSA, bovine serum albumin) were injected into the flow cell and exposed to X-ray radiation (gallium Kα emission of 9.2 keV) produced by a MetalJet D2 microfocus generator (Excillum AB, Kista, Sweden). For each sample, 10 independent frames (exposure time 600 s per frame) were recorded using a PILATUS3 1 M detector (DECTRIS Ltd., Baden-Daettwil, Switzerland). Scattering data for ggNucb2 and hsNucb2 were integrated using Foxtrot [114] (link) and processed using the SAS data analysis module from the ATSAS package [115] (link).
Xeuss 2.0 saxs waxs system
Manufactured by Xenocs
Sourced in France
The Xeuss 2.0 SAXS/WAXS system is a laboratory instrument designed for small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) analysis. It provides high-resolution data collection and processing capabilities for the characterization of various materials, including polymers, nanomaterials, and biological samples.
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Lab products found in correlation
Pilatus3 1m detector, by Dectris (2 mentions)
Genix3d cu uld, by Dectris (2 mentions)
Pilatus 300k, by Dectris (2 mentions)
Pilatus 3r 1m detector, by Dectris (1 mentions)
Thms600 hot stage, by Linkam (1 mentions)
Lambda 950 spectrophotometer, by PerkinElmer (1 mentions)
R2658p pmt, by Hamamatsu Photonics (1 mentions)
Pilatus3 r 300k detector, by Dectris (1 mentions)
9 protocols using xeuss 2.0 saxs waxs system
1
SAXS Analysis of Nucb2 Proteins
2
X-ray Diffraction Analysis of PrP58–93 Fibrils
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X-ray fiber diffraction experiment of PrP58–93 fibrils was conducted on XEUSS 2.0 SAXS/WAXS system (XENOCS, Sassenage, France), using Ga Kα radiation from a MetalJet microfocus generator with a liquid-metal jet anode (Excillum AB, Kista, Sweden) and a Pilatus 3R 1M detector (DECTRIS Ltd. Baden-Daettwil, Switzerland). The sample-to-detector distance was calibrated by the use of silver behenate (reference q001 = 1.070 nm−1). 500 μl of 0.4 mM PrP58–93 peptide prepared in 50 mM HEPES, pH 7.4, 100 mM NaCl with 1.6 mM ZnCl2 was incubated at room temperature for 24 h. After incubation time, the aggregated deposits, visible on the walls of the Eppendorf test tube, were transferred into thin-walled borosilicate glass and the data were collected.
3
SAXS and WAXS Characterization Protocol
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SAXS (small-angle X-ray scattering) and WAXS (wide-angle X-ray scattering) measurements were measured on a Xeuss 2.0 SAXS/WAXS system (Xenocs SA, France). Cu Kα X-ray source (GeniX3D Cu ULD), generated at 50 kV and 0.6 mA, was utilized to produce X-ray radiation with a wavelength of 1.5418 Å. A semiconductor detector (Pilatus 300 K, DECTRIS, Swiss) with a resolution of 487 × 619 pixels (pixel size = 172 × 172 μm2) was used to collect the scattering signals. For two dimensional (2D) SAXS, the sample-to-detector distance was 1196.26 mm, which was determined by a Silver Behenate (AgC22H43O2) standard. Each WAXS pattern was collected with an exposure time of 6 min. with 2θ range of 1.8-30 degree. The one-dimensional intensity profiles were integrated from background corrected 2D WAXS patterns (Supplementary Fig. 28 ). Beamstop: D = 5 mm.
4
WAXS analysis of electrospun mats
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WAXS measurements of electrospun mats were performed at the University of Delaware on a Xeuss 2.0 SAXS/WAXS system (Xenocs, Sassenage, France) with the X-ray source operated at a voltage of 50 kV and a current of 0.6 mA. Monochromatic X-rays with a wavelength of 1.542 Å (Cu K radiation) were used to irradiate the samples at a sample-to-detector distance of 72 mm as determined by calibration using a silver behenate standard. Scattering patterns were recorded on a Pilatus detector (comprised of three panels) with a total number of pixels using 1-h exposure times.
5
SAXS Measurements with Xeuss 2.0 System
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Small-angle X-ray scattering (SAXS) measurements were performed using a Xeuss 2.0 SAXS/WAXS system (Xenocs, France) with pinhole collimation for point focus geometry [18 (link)]. The SAXS camera was equipped with a Hi-Star 2D detector. The optics and sample chamber were used under vacuum conditions to minimize air scatter. The liquid samples were placed in 0.5-mm glass capillaries, and measurements were performed in a vacuum at 25 °C with an exposure time of 0.5 h.
6
SAXS Analysis of 3D Protein Assemblies
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Small angle X-ray scattering measurements were experimented using a Xeuss2.0 SAXS/ WAXS system (Xenocs, France). Blank sample with no protein was collected before experiment sample and a standard sample (silver behenate) were used to calibrate the scattering length vector q. Experimental sample was obtained from sediment after centrifuging when 3D assemblies formed. The magnitude of the scattering vector q is given by q = 4 π sin θ/λ, where 2θ is the scattering angle. Sample thickness was approximately 1.5 mm. The simulated scattering patterns were obtained with Scatter (version 2.5).
7
In Situ WAXS Characterization of Thermal Transitions
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WAXS measurements
were performed on a Xeuss 2.0 SAXS/WAXS system (Xenocs SA, France).
X-ray radiation (wavelength = 1.5418 Å) was produced by means
of the Cu Kα radiation generator (GeniX3D Cu ULD) at 50 kV and
0.6 mA. Scattered signals were collected by a semiconductor detector
(Pilatus 300 K, DECTRIS, Swiss) with a resolution of 487 × 619
pixels (pixel size 172 × 172 μm2). Each room
temperature WAXS pattern was obtained with 20 min exposure time. The
one-dimensional intensity profiles were integrated from background
corrected 2D WAXS patterns with an azimuthal angle range of 0–90°.
Transmission geometry was adopted for in situ measurements.
The temperature was controlled by a Linkam THMS600 hot stage (Linkam
Scientific Instruments, UK). Heating and cooling rates for the measurement
were set at 20 °C/min. Specimens were held for 1 min at the selected
temperature to stabilize the temperature, and then WAXS were obtained
with 5 min exposure times. The thermal protocol consisted of four
heating steps (200, 215, 235, and 260 °C) and nine cooling steps
(250, 240, 230, 220, 210, 200, 190, 180, and 150 °C). WAXS patterns
were collected at room temperature (∼30 °C) before the
beginning and after the completion of the thermal protocol to evaluate
structural changes which could occur while keeping the material at
high temperatures for long times.
were performed on a Xeuss 2.0 SAXS/WAXS system (Xenocs SA, France).
X-ray radiation (wavelength = 1.5418 Å) was produced by means
of the Cu Kα radiation generator (GeniX3D Cu ULD) at 50 kV and
0.6 mA. Scattered signals were collected by a semiconductor detector
(Pilatus 300 K, DECTRIS, Swiss) with a resolution of 487 × 619
pixels (pixel size 172 × 172 μm2). Each room
temperature WAXS pattern was obtained with 20 min exposure time. The
one-dimensional intensity profiles were integrated from background
corrected 2D WAXS patterns with an azimuthal angle range of 0–90°.
Transmission geometry was adopted for in situ measurements.
The temperature was controlled by a Linkam THMS600 hot stage (Linkam
Scientific Instruments, UK). Heating and cooling rates for the measurement
were set at 20 °C/min. Specimens were held for 1 min at the selected
temperature to stabilize the temperature, and then WAXS were obtained
with 5 min exposure times. The thermal protocol consisted of four
heating steps (200, 215, 235, and 260 °C) and nine cooling steps
(250, 240, 230, 220, 210, 200, 190, 180, and 150 °C). WAXS patterns
were collected at room temperature (∼30 °C) before the
beginning and after the completion of the thermal protocol to evaluate
structural changes which could occur while keeping the material at
high temperatures for long times.
8
FTO Homodimer SAXS Analysis
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BESFTO (26 µM) was incubated for 5 min in SAXS1 buffer and then loaded into a low noise liquid sample cell (Xenocs, Grenoble, France). Samples were measured using an XEUSS 2.0 SAXS/WAXS system (Xenocs, Grenoble, France) with X-ray radiation (gallium Kα emission of 9.2 keV) produced by a MetalJet D2 microfocus generator (Excillum AB, Kista, Sweden). For each sample, 5-10 independent frames (exposition time 600 s per frame) were recorded using the PILATUS3 1M detector (DECTRIS Ltd., Baden-Daettwil, Switzerland). Scattering data were integrated using Foxtrot [55 (link)]. Data were analyzed using SCÅTTER software (http://www.bioisis.net/ ; accessed on 1 February 2020) and the PRIMUS program from the ATSAS package was used for curve fitting. Structural parameters (Rg Guinier and Rg p(r)) were estimated using the GNOM program from the ATSAS package [56 (link)]. Low resolution bead models were generated using the DAMMIN program [57 (link)]. The ensemble of FTO homodimer structures was obtained with the use of a model generated by the SymmDock server (http://bioinfo3d.cs.tau.ac.il/SymmDock/ ; accessed on 1 February 2020) [58 (link)].
9
Advanced Characterization of Thin Films
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X-ray reflectivity was measured on a diffractometer (3303TT, GE) using Cu Kα radiation (λ = 1.5406 Å) and a 1D detector (Meteor 1D, XRD Eigenmann). Grazing-incidence X-ray diffraction and the reciprocal space maps were measured at beamline SixS at Soleil, Gif-sur-Yvette, France, using a wavelength of λ = 0.9538 Å and on a Xeuss 2.0 SAXS/WAXS system (Xenocs) with a Dectris Pilatus3R 300-K detector (λ = 1.5406 Å). The sample was kept in vacuum during the measurements to avoid beam damage.
UV–vis transmission spectra were measured using a Perkin Elmer Lambda 950 spectrophotometer. Photoluminescence-excitation spectroscopy was performed on a Horiba Fluorolog-3 DF spectrofluorimeter using a 450-W xenon lamp for excitation and a Hamamatsu R2658P PMT to monitor the emission.
UV–vis transmission spectra were measured using a Perkin Elmer Lambda 950 spectrophotometer. Photoluminescence-excitation spectroscopy was performed on a Horiba Fluorolog-3 DF spectrofluorimeter using a 450-W xenon lamp for excitation and a Hamamatsu R2658P PMT to monitor the emission.
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