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Synchrotron SAXS data from solutions of the complement C1r CCP2-SP domains in 10 mM HEPES, pH 7.3, 140 mM NaCl, were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS; (Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2.1 m and at a wavelength of λ = 0.1127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 55.00 μl sample at 2 mg/ml was injected at a 0.50 ml/min flow rate onto a Shodex KW-802.5 column at 20°C. 3 second X-ray exposures were collected continuously during the entire 30 min elution.The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted from the appropriate sample peak frames.
The beam line is equipped with an online Agilent 1260 Infinity HPLC system with a series of UV at 280 and 260 nm, multi-angle light scattering (MALS), quasi-elastic light scattering (QELS), and refractometer detectors. MALS experiments were performed using an 18-angle DAWN HELEOS II light scattering detector connected in tandem to an Optilab refractive index concentration detector (Wyatt Technology).
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s, nm-1
