Clarified harvests from each culture were purified using 2 ml of Galanthus nivalis Lectin (GNL) agarose (Catalog# AL1243–5, Vector Laboratories, Burlingame, California USA) packed in glass columns (Omnifit glass column, 006BCC-10–10-AF, 10 mm/ 100 mm, Kinesis, IL, USA). Each column was equilibrated with 20 mM Tris pH 8.0, 500 mM sodium chloride, 0.1% Triton™ X-100 at 200 cm/h for 7.5 column volumes. Clarified culture media was loaded at 100 cm/h. The column was then washed with 20 mM Tris pH 8.0, 500 mM sodium chloride, 0.1% Triton™ X-100 followed by 50 mM Tris pH 8.0, 150 mM sodium chloride at 200 cm/h for 7 column volumes. Elution was performed using 50 mM Tris pH 8.0, 150 mM sodium chloride, and 0.5 M mannopyranoside at 75 cm/h. Fractions with the highest absorbance were pooled and filtered using 0.8/0.2 µm Acrodisc PF Syringe Filters (Catalog #P4187, Pall Corporation, Baltimore, Maryland USA).
Acrodisc pf syringe filters
Manufactured by Pall Corporation
Sourced in United States
The Acrodisc PF Syringe Filters are used for the filtration of samples prior to analysis. The filters are designed to remove particulates and clarify samples, ensuring the integrity of analytical results.
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Lab products found in correlation
2 protocols using acrodisc pf syringe filters
1
Galanthus nivalis Lectin Purification
2
Aqueous Solubility Determination of HHFA, HHFX, HHFG, HHFMan
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For estimating the aqueous solubility of the samples, 1% wt, 0.1% wt and 0.01% wt solutions were prepared in deionized or salted aqueous media. For homogenization, the samples were vigorously stirred with a magnetic stirrer in a water bath at 50 °C for approximately 3 hours, and then left at 50 °C during 15 hours in an oven.
For the 1% wt and 0.1% wt samples, the solubility was visually inspected at 50 °C, and then after 15 hours at room temperature and at 83 °C. For the 0.01% wt samples, the solubility was visually inspected at room temperature, and after 15 hours at 65.5 °C and 83 °C.
For determining aqueous solubility limits of HHFA, HHFX, HHFG and HHFMan samples, the 0.01% wt solutions were filtered on 0.2 μm pore size syringe filter (Pall Corporation, Acrodisc PF syringe filters, diameter 0.8 cm/0.2 μm) two consecutive times at room temperature. The filtrate was dosed by HPLC/MS, thanks to a calibration line established beforehand on a 5–100 ppm scale.
For the 1% wt and 0.1% wt samples, the solubility was visually inspected at 50 °C, and then after 15 hours at room temperature and at 83 °C. For the 0.01% wt samples, the solubility was visually inspected at room temperature, and after 15 hours at 65.5 °C and 83 °C.
For determining aqueous solubility limits of HHFA, HHFX, HHFG and HHFMan samples, the 0.01% wt solutions were filtered on 0.2 μm pore size syringe filter (Pall Corporation, Acrodisc PF syringe filters, diameter 0.8 cm/0.2 μm) two consecutive times at room temperature. The filtrate was dosed by HPLC/MS, thanks to a calibration line established beforehand on a 5–100 ppm scale.
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