Hyaluronan binding by soluble sTMEM2 (SEC)

Size exclusion chromatography of 1 mg/ml transmembrane protein 2 ectodomain (sTMEM2) either alone or pre-incubated for 30 minutes with 200 μg/ml high-molecular-weight hyaluronan (HMW-HA, >950 kDa, R&D Systems). The running buffer was 50 mM HEPES, 125 mM NaCl, 2 mM CaCl₂, pH 7.5.

本实验采用尺寸排阻色谱法（Size Exclusion Chromatography），对两类样本开展检测：其一为浓度1 mg/ml的单独可溶性跨膜蛋白2胞外域（sTMEM2）；其二为该蛋白与200 μg/ml高分子量透明质酸（HMW-HA，>950 kDa，R&D Systems）预孵育30分钟后的混合体系。所用运行缓冲液为含50 mM 4-羟乙基哌嗪乙磺酸（HEPES）、125 mM氯化钠、2 mM氯化钙且pH值为7.5的缓冲溶液。