Ultimate iso 3100sd pump

High Performance Size Exclusion Chromatography was performed using an Ultimate iso-3100 SD pump with a WPS-3000 sampler (Dionex, Sunnyvale, CA, USA) connected to an RI-101 refractive index detector (Shodex, Showa Denko K.K., Tokyo, Japan). One hundred microliters of three times diluted reaction mixtures were loaded on a Shodex SB-806 HQ GPC column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 mm × 6 mm) (Showa Denko K.K., Tokyo, Japan). Elution was performed with 100 mM sodium acetate pH 6 at a flow rate of 0.5 mL/min at room temperature. Pullunan standards were used as references.

The size of the fucoidan substrates and Mef1-hydrolyzed products were determined by HP-SEC using an Ultimate iso-3100SD pump with a WPS-3000 sampler (Dionex, Sunnyvale, California, USA) connected to an RI-101 refractive-index (RI) detector (Shodex, Showa Denko K.K., Tokyo, Japan). Samples were separated on a Shodex SB-806 HQ GPC column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 × 6 mm) with elution using 100 mM sodium acetate buffer pH 6 at a flow rate of 0.5 ml min⁻¹ at 40°C (Trang et al., 2022 ▸ ). Pullulans of 1, 5, 12, 110, 400 and 800 kDa (Sigma–Aldrich, Steinheim, Germany) were used as standards.

The size of the fucoidan substrates and Mef1-hydrolyzed products were determined by HP-SEC using an Ultimate iso-3100SD pump with a WPS-3000 sampler (Dionex, Sunnyvale, California, USA) connected to an RI-101 refractive-index (RI) detector (Shodex, Showa Denko K.K., Tokyo, Japan). Samples were separated on a Shodex SB-806 HQ GPC column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 × 6 mm) with elution using 100 mM sodium acetate buffer pH 6 at a flow rate of 0.5 ml min⁻¹ at 40°C (Trang et al., 2022 ▸ ). Pullulans of 1, 5, 12, 110, 400 and 800 kDa (Sigma–Aldrich, Steinheim, Germany) were used as standards.

Freeze-dried citrus peel xyloglucan was dissolved in 100 mM acetate buffer (pH 4.6) to a concentration corresponding to 8 mM bound Fuc (approximately 0.1 g/L). To investigate the effect of substrate depolymerization by xyloglucanase treatment, the extracted xyloglucan was treated with a GH5 Paenibacillus sp. xyloglucan-specific endo-β-1,4-glucanase (XEG; EC 3.2.1.151; Megazyme, Wicklow, Ireland) using 100 U/g substrate at 40 °C for 1 h. The xyloglucanase was heat-inactivated for 10 min at 99 °C. The untreated, extracted xyloglucan was subjected to the same heat treatment without enzyme addition. Size distributions of the two xyloglucan preparations were determined by size exclusion chromatography (SEC) as described previously [29 (link)] using a Shodex SB-806 HQ GPC column (300 mm × 8 mm) equipped with a Shodex SB-G guard column (50 mm × 6 mm; Showa Denko K.K., Tokyo, Japan), an UltiMate iso-3100 SD pump (Dionex, ThermoFisher Scientific, Waltham, MA, USA), and an RI-101 refractive index detector (Showa Denko K.K., Tokyo, Japan). Elution took place with a 100 mM sodium acetate eluent at a flow rate of 0.5 mL/min at 40 °C. Pullulan standards were used as the reference for molecular weight.

High Performance Size Exclusion Chromatography was performed using an Ultimate iso-3100 SD pump with a WPS-3000 sampler (Dionex, Sunnyvale, CA, USA) connected to an RI-101 Shodex refractive index detector (Showa Denko K.K, Tokyo, Japan). 100 µL of reaction samples diluted three times was loaded on a Shodex SB-806 HQ GPC column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 × 6 mm) (Showa Denko K.K, Tokyo, Japan). Elution was performed using 100 mM sodium acetate pH 6 at a flow rate of 0.5 mL·min⁻¹ at room temperature. Pullulan of 800, 400, 110, 12, and 5 kDa (Sigma-Aldrich, Steinheim, Germany) were used as standards.