SPR analysis was performed using the BIAcore T200 model (GE Healthcare) at room temperature with PBSTT buffer (1X PBS, 0.05% Triton X-100, 0.05% Tween-20) containing 5% dimethyl sulfoxide (DMSO; Sigma-Aldrich; Merck KGaA). The pH scouting for GP130 (Sino Biological, China, or ANRT, Korea) and IL-6Rα (PeproTech) immobilisation was performed in 10 mM acetate buffer at pH 4.0, 4.5, 5.0, and 5.5. GP130 and IL-6Rα were immobilised on a CM5 chip to 2,000 and 1,000 response units (RU) with standard amine coupling at pH 4.5. Bazedoxifene was injected into the GP130 and IL-6Rα-immobilised flow cell at concentrations of 50, 100, 125, 150, 175, and 200 µM with a flow rate of 20 µl/min for 250 sec and allowed to dissociate for 600 sec. The T-200 BIAevaluation software v3.1 (GE Healthcare) was used to subtract the references and determine steady-state KD. Between the sample series, a solvent correction cycle was run to adjust for referencing errors caused by refractive index mismatches between the running buffer and samples (24 (link)).
Il 6rα
Manufactured by GE Healthcare
Sourced in United States
IL-6Rα is a lab equipment product that functions as a receptor for the cytokine interleukin-6. It is used in research and clinical applications to study the role of interleukin-6 signaling in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using il 6rα
1
SPR Analysis of Bazedoxifene Binding
2
Surface Plasmon Resonance Binding Assay
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The surface plasmon resonance (SPR) assay was measured using the BIAcore T200 model (GE Healthcare, Chicago, IL, USA) with HBS-EP buffer (1×HBS-EP, GE Healthcare, Chicago, IL, USA) containing 5% dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA). The pH scouting for IL-6 (PeproTech, Rocky Hill, NJ, USA), IL-6Rα (PeproTech, Rocky Hill, NJ, USA), and gp130 (Sino Biological, Beijing, China, or ANRT, Daejeon, Korea) immobilization was performed in 10 mM acetate buffer at pH 4.5. IL-6, IL-6Rα, and gp130 were immobilized on a CM5 chip at 750, 1200, and 2000 response units (RU). Butein was injected into the IL-6, IL-6Rα and gp130-immobilized flow cell at concentrations of 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 μM with a 25 μL/min flow rate for 150 s and allowed to dissociate for 300 s. Steady-state KD was determined via the T-200 BIAevaluation software (GE Healthcare, Chicago, IL, USA).
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