Dextran t 500

Manufactured by Carl Roth

Sourced in Germany

Dextran T 500 is a high molecular weight polysaccharide derived from the bacteria Leuconostoc mesenteroides. It is commonly used in laboratory applications as a density gradient medium for the separation and purification of cells, organelles, and macromolecules.

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Lab products found in correlation

Pancoll solution, by PAN Biotech (1 mentions) Riboflavin, by Serva Electrophoresis (1 mentions) Dextran, by Carl Roth (1 mentions) Riboflavin, by Carl Roth (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions) Macsxpress human neutrophil isolation kit, by Miltenyi Biotec (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Cd14 microbead, by Miltenyi Biotec (1 mentions) Ficoll plaque plus, by GE Healthcare (1 mentions) Hank s balanced salt solution, by Lonza (1 mentions)

4 protocols using dextran t 500

Isolation of Human Neutrophils from Buffy Coats

Cited 1 time

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Human neutrophils were isolated from buffy coats as previously described (57 (link)). In brief, dextran T 500 (3%) (Carl Roth GmbH, Germany) (3% dextran T 500 in 0.9% NaCl, at 4:1) was added to the buffy coats in 15-mL tubes, and erythrocytes were sedimented at gravity for 20 min. The white blood cell layer was removed carefully, and cells were pelleted by centrifugation at 200 × g for 20 min. The cell pellet was resuspended in 12 mL of Hanks’ balanced salt solution (HBSS; Lonza Cologne GmbH, Germany), layered on top of 3 mL Ficoll-plaque Plus (GE Healthcare, Germany), and centrifuged at 400 × g for 20 min. Next, residual erythrocytes were lysed in lysis buffer (150 mM NH₄Cl, 10 mM KHCO₃, and 10 mM EDTA [pH 7.4]). The neutrophils were washed (200 × g for 10 min), suspended in HBSS, and used immediately.

Dasari P., Nordengrün M., Vilhena C., Steil L., Abdurrahman G., Surmann K., Dhople V., Lahrberg J., Bachert C., Skerka C., Völker U., Bröker B.M, & Zipfel P.F. (2022). The Protease SplB of Staphylococcus aureus Targets Host Complement Components and Inhibits Complement-Mediated Bacterial Opsonophagocytosis. Journal of Bacteriology, 204(1), e00184-21.

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Neutrophil Isolation from Whole Blood

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Human neutrophils were isolated from whole blood by a 3% dextran sedimentation step (Dextran T500, Carl Roth GmbH, Germany) followed by centrifugation over a 1.077 g/ml Pancoll solution (PAN-Biotech). Cell viability was evaluated by trypan blue exclusion, and purity (> 95%) was verified by flow cytometry. After isolation, neutrophils were resuspended in HBSS +/+, quantified, and used for experiments as indicated.

Letsiou E., Teixeira Alves L.G., Fatykhova D., Felten M., Mitchell T.J., Müller-Redetzky H.C., Hocke A.C, & Witzenrath M. (2021). Microvesicles released from pneumolysin-stimulated lung epithelial cells carry mitochondrial cargo and suppress neutrophil oxidative burst. Scientific Reports, 11, 9529.

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Riboflavin-Dextran UV Tissue Treatment

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Pericardial tissue was treated with 260 μM riboflavin (Serva, Germany) and 2% dextran T500 (Carl Roth, Germany) for at least 24 h. To prevent drying, samples were placed on filter paper immersed in riboflavin/dextran solution and irradiated with 570 mJ/cm² UV (320–480 nm) within 30 min.

Walker S., Schönfelder J., Tugtekin S.M., Wetzel C., Hacker M.C, & Schulz-Siegmund M. (2018). Stabilization and Sterilization of Pericardial Scaffolds by Ultraviolet and Low-Energy Electron Irradiation. Tissue Engineering. Part C, Methods, 24(12), 717-729.

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Isolation and Culture of Neutrophils and Monocytes

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Neutrophils were isolated as described ^{38 (link)}. The procedure included a density gradient centrifugation step of whole blood on Ficoll-Hypaque (Pharmacia Diagnostics, Uppsala, Sweden), dextran sedimentation (4% dextran T 500, Carl ROTH, Germany), red cell osmotic lysis of the granulocyte pellet, and a final immunomagnetic negative selection step using the human MACSxpress^® Neutrophil Isolation Kit (Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany) and resulted repeatedly in >99% pure CD16⁺CD66b⁺CCR3^-HLA-DR^-CD19^-CD3^-CD14^- cells. Neutrophils (1x10⁶/ml) were cultured in RPMI 1640 (Sigma Aldrich, Darmstadt, Germany) supplemented with 10% autologous plasma alone or stimulated with GM-CSF (100 pg/ml), IFN-γ (10 ng/ml) and IL-3 (30 ng/ml; all from Peprotech, Rocky Hill, USA) as described ^{28 (link)}. Monocytes (>98% CD14⁺CCR3^-CD19^-CD3^-) were obtained by using CD14 MicroBeads (Miltenyi Biotec) according to the manufacturer´s instructions.

Polak D., Hafner C., Briza P., Kitzmüller C., Elbe-Bürger A., Samadi N., Gschwandtner M., Pfützner W., Zlabinger G.J., Jahn-Schmid B, & Bohle B. (2018). A novel role for neutrophils in IgE-mediated allergy: evidence for antigen-presentation in late-phase reactions. The Journal of allergy and clinical immunology, 143(3), 1143-1152.e4.

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