Size exclusion chromatography–multiangle light scattering

The purified CgHspB1 was analyzed by size exclusion chromatography–multiangle light scattering (SEC‐MALS) on a TSKgel G3000XL column (Tosoh) connected to a multiangle light‐scattering detector (MINI DAWN; Wyatt Technology, Santa Barbara, CA, USA) and a differential refractive index detector (Shodex RI‐101; Showa Denko) with an HPLC system, PU‐980i (JASCO), as described previously 31. A 100‐µL aliquot of sample was injected into the column and eluted with buffer B at 1.0 mL·min⁻¹. The molecular weight and protein concentration were determined according to the instructional manual (Wyatt Technology).