Lymphopure™ is a ready-to-use filtered and endotoxin tested solution for the isolation of mononuclear cells (MNCs) from a cell suspension. The solution is formulated at a density that allows separation of MNCs from other cells in blood such as granulocytes and erythrocytes. Granulocytes and erythrocytes will sediment after centrifugation.
Formulation: Density gradient medium for the isolation of mononuclear cells formulated at a density of 1.077 ± 0.001 g/mL, and osmolality of 290 ± 15 mOsm.
Storage & Handling: Upon receipt, store at room temperature protected from light.
Application: Cell Separation - Quality tested
Recommended Usage: Watch a video protocol.
- Collect blood into a tube containing anti-coagulant (EDTA, heparin)
- Dilute the blood with an equal volume of phosphate-buffered saline or similar suitable medium.
- Mix Lymphopure™ by inverting the bottle several times.
- Aliquot half the amount of Lymphopure™ into a tube. For every ml of diluted blood, add 0.5 ml of Lymphopure™.
- Carefully layer the diluted blood on top of Lymphopure™. Avoid mixing blood with Lymphopure™.
- Centrifuge at 800 x g for 20 minutes at room temperature (15 - 25°C) without braking. If the blood has been stored for more than 2 hours, centrifuge for 30 minutes.
- After centrifugation, mononuclear cells (MNCs) form a defined cell layer at the plasma: Lymphopure™ interface.
- Collect the MNC layer at the interface without disturbing the other fractions.
- Wash MNCs with medium.
Application References: (PubMed link indicates BioLegend citation)
- Böyum A. 1968. Scand. J. Clin. Lab. Invest. Suppl. 97:9-29.
- Böyum A. 1968. Scand. J. Clin. Lab. Invest. Suppl. 97:77-89.
- Böyum A. 1964. Nature. 204:793-4.
- Thorsby E, Bratile A. 1970. Histocompatibility Testing 1970,ed. P.I. Terasaki. 655-6.
- Ting A, Morris PJ. 1971. Vox. Sang. 20(6)561-3.
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