The glycans from the second and third extractions were pooled (~20 mg) and applied to a DEAE-cellulose column (GE Healthcare Life Sciences, United Kingdom), linked to a FPLC Äkta Prime system (GE Healthcare Life Sciences, United Kingdom). The column was equilibrated with 0.5 M sodium acetate (pH 6.0) and washed with 50 mL of the same buffer. The column was developed by an increasing concentration of NaCl (0.5→1.0 M) in the same buffer. The flow rate of the column was 8.0 mL h−1, and fractions of 1.5 mL were collected and analyzed by metachromasia using 1,9-dimethylmethylene blue [28 (link)]. The NaCl concentration was estimated by conductivity. Fractions under the peaks were pooled, dialyzed against distilled water, lyophilized, and analyzed by agarose gel electrophoresis.
Deae cellulose column
Manufactured by GE Healthcare
Sourced in United Kingdom
DEAE-cellulose column is a type of ion-exchange chromatography column. It is used for the separation and purification of biomolecules, such as proteins, nucleic acids, and other charged molecules, based on their differences in charge interactions with the DEAE (diethylaminoethyl) functional groups present on the cellulose matrix.
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Lab products found in correlation
2 protocols using deae cellulose column
1
Glycan Purification via DEAE-Cellulose Chromatography
2
Broccoli Extract Purification
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Two volumes of PCI (phenol/chloroform/isoamyl alcohol) was added to the hot water extract of broccoli, and the aqueous layer was collected after centrifugation (8,000 rpm, 5 minutes). Two volumes of ethanol was added to the aqueous layer, and the precipitate formed by centrifugation (8,000 rpm, 10 min) was dissolved in 10 mM Tris-HCl (pH 7.9) buffer, and loaded to a DEAE cellulose column (GE Healthcare, bed volume = 20 mL) equilibrated by the same buffer, and we collected the flow-through fraction.
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