One hundred pairs of female An. gambiae SGH were dissected under the microscope, sonicated extensively, and centrifuged for 10 min at 13,000×g. The supernatant (∼100 µl) was collected and loaded in a Superdex G-75 10/300 GL gel-filtration column (GE healthcare Biosciences) equilibrated in PBS pH 7.4 containing 0.1% PEG 4000. Elution was carried out at 0.5 ml/min, with 0.5 ml fractions. Addition of PEG was critical in order to recover the hemagglutinating activity of the SGH. The fractions were subsequently tested for hemagglutinating activity as above. Control experiments demonstrated that the presence of PEG 0.1% did not affect the lectin properties of the SGH. In order to estimate the correct molecular weight of the candidates, the column was calibrated with Conalbumin (75 kDa), BSA (66 kDa), Ovalbumin (40 kDa), and Approtinin (6.5 kDa).
Superdex g 75 10 300 gl gel filtration column
Manufactured by GE Healthcare
Sourced in Sweden
The Superdex G-75 10/300 GL gel-filtration column is a laboratory equipment used for the separation and purification of biomolecules based on their size and molecular weight. It is a pre-packed column designed for use with fast protein liquid chromatography (FPLC) systems, providing efficient and reproducible separations of proteins, peptides, and other macromolecules.
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Lab products found in correlation
2 protocols using superdex g 75 10 300 gl gel filtration column
1
Purification of Mosquito Salivary Lectin
2
Extraction and Purification of Tartary Buckwheat Compounds
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Tartary buckwheat seeds were obtained from Shanxi Academy of Agricultural Sciences, China. A Resource™ Q (6.4 x 30 mm) anion exchange column and Superdex™ G75 10/300 GL gel filtration column were purchased from GE Healthcare (Uppsala, Sweden), RhIL-4 and GM-CSF were purchased from R&D Systems (Minneapolis, MN, USA), and fetal bovine serum (FBS) and cell medium RPMI1640 were sourced from Sigma-Aldrich Co., Ltd., (St. Louis, MO, USA). Mouse anti-human CD83-PE monoclonal antibody, mouse anti-human CD40-FITC monoclonal antibody, mouse anti-human CD1a-FITC monoclonal antibody, and mouse anti-human HLA-DR monoclonal antibody were all products of Beckman Coulter Inc. (Brea, CA, USA). All solvents used in high performance liquid chromatography were of chromatography grade, and all other reagents were of the highest purity available.
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