Free HisBP after TEV cleavage to remove hexahistidine tags, followed by reverse His-Trap and SEC column purification. Note N-terminal glycine before the Uniprot sequence due to TEV tag.
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1. 样品、组分与实验条件 Sample & Experiment
样品 1 · Histidine-binding periplasmic protein (HisBP), apo-form
Synchrotron SAXS data from solutions of HisBP, apo-form, in 100 mM NaCl, 20 mM NaPO4, 0.5 mM TCEP, pH 7.4 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.09919 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.25 mg/ml was measured at 20°C. 12 successive 2 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
相关论文
标题
Structure-based screening of binding affinities via small-angle X-ray scattering
European Molecular Biology Laboratory (EMBL) - Heidelberg
experiment.contributor[0].affiliation[0].webpage
https://www.embl.de/
experiment.contributor[0].contributor_name
Po-chia
experiment.contributor[0].contributor_surname
Chen
experiment.contributor[0].orcid
null
experiment.concentration_method
null
experiment.concentration_unit
null
experiment.date
2018-09-10
experiment.storage_temperature
null
experiment.cell_temperature
20.0
experiment.exposure_time
2.0
experiment.number_of_frames
12
experiment.wavelength
0.09919
experiment.sample_detector_distance
2.849
experiment.concentration_min
null
experiment.concentration_max
4.25
experiment.sample_volume
null
experiment.flow_rate
null
experiment.s_min
0.031
experiment.s_max
4.946
experiment.total_exposure_time
null
experiment.seccolumn
null
fits
[]
estimated_volume_method
null
pddf_software
ATSAS GNOM
pddf_software_version
5.0
i0_calibration_standard
null
description
Free HisBP after TEV cleavage to remove hexahistidine tags, followed by reverse His-Trap and SEC column purification. Note N-terminal glycine before the Uniprot sequence due to TEV tag.
experiment_description
Synchrotron SAXS data from solutions of HisBP, apo-form, in 100 mM NaCl, 20 mM NaPO4, 0.5 mM TCEP, pH 7.4 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.09919 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.25 mg/ml was measured at 20°C. 12 successive 2 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.