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| Product Name | Lymphocyte Separation Medium 1.077 |
| Catalog No. | RESK-0255 |
| Description | Lymphocyte Separation Medium (LSM) is a filter-sterilized, ready-to-use separation medium for the in vitro isolation of lymphocytes from human peripheral blood, and can also be used to isolate lymphocytes from other tissues, including umbilical cord blood and bone marrow. Each 100 mL contains 6.2 g polysucrose and 9.4 g sodium diatrizoate. The density is 1.0770–1.0800 g/mL at 20 °C. |
| Intended Use | In vitro isolation of lymphocytes (mononuclear cells) from human peripheral blood; also applicable to lymphocyte isolation from umbilical cord blood and bone marrow cells. For research use only. |
| Principle / Technology | Defibrinated or heparin-treated human blood is diluted 1:1 with physiological saline or balanced salt solution, layered onto the separation medium, and centrifuged at low speed for 30 min. During centrifugation, differential migration produces distinct cell layers: erythrocytes and polymorphonuclear granulocytes migrate through the gradient to the bottom of the tube, while lymphocytes and other mononuclear cells (monocytes and platelets), due to their lower density, are distributed at the interface between the plasma and the separation medium. Lymphocytes are recovered by aspirating the interface layer and further washed to remove platelets, medium and plasma. |
| Detection Method | Density gradient centrifugation; cells are recovered from the plasma-medium interface. |
| Sample Type | Human peripheral blood (defibrinated or anticoagulant-treated); umbilical cord blood; bone marrow cells |
| Performance Range / Specifications | Density: 1.0770–1.0800 g/mL at 20 °C; sterile, ready-to-use solution. |
| Reaction Conditions / Protocol | (For human blood treated with defibrination or anticoagulants; for blood from other species or tissues, the procedure should be modified accordingly) 1. Gently invert the bottle to mix the medium thoroughly. 2. Aseptically transfer 3 mL of the medium into a 15 mL centrifuge tube. 3. Mix 2 mL of defibrinated or heparinized blood with 2 mL of physiological saline. 4. Carefully layer the diluted blood onto 3 mL of the medium (at room temperature) in the 15 mL centrifuge tube to form a distinct interface; do not mix the diluted blood into the medium. 5. Centrifuge at 400 × g for 15–30 min at room temperature. Centrifugation pellets erythrocytes and polymorphonuclear leukocytes while a band of mononuclear lymphocytes forms on top of the medium (from top to bottom: plasma layer – mononuclear cell layer – separation medium layer – RBC pellet). 6. Aspirate the plasma to within 2–3 mm above the lymphocyte layer. 7. Transfer the lymphocyte layer together with about half of the underlying medium layer to another centrifuge tube. Add an equal volume of balanced salt buffer and centrifuge for 10 min at room temperature (18–25 °C) at a speed that pellets the cells without damaging them, e.g., 160–260 × g (this washes away the medium and reduces the platelet percentage). 8. Wash the cells again with balanced salt buffer and finally resuspend in an appropriate medium of the user's choice. |
| Components / Formulation | Each 100 mL contains 6.2 g polysucrose and 9.4 g sodium diatrizoate; filter-sterilized, ready-to-use solution. |
| Storage Conditions | Store at 15–30 °C. |
| Shelf Life | 2 years |
| Package Specifications | 100 mL 5 × 100 mL |
| Product Form | Sterile liquid, ready-to-use |
| Quality Control | Filter-sterilized; density 1.0770–1.0800 g/mL at 20 °C |
| Concentration | 6.2 g polysucrose and 9.4 g sodium diatrizoate per 100 mL |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | 2 years when stored at 15–30 °C. |
| Regulatory / Compliance | For research use only. |
| Recommended Buffer System | Physiological saline or balanced salt solution for blood dilution and cell washing. |
| Application Notes / Precautions | 1) For your safety and health, wear a lab coat and disposable gloves during operation. 2) The physiological saline used to dilute defibrinated or heparinized blood must be sterile. 3) To preserve lymphocyte viability, separation should be performed as soon as possible after blood collection. The recovered cell layer is actually the mononuclear cell layer, including lymphocytes and monocytes. 4) This product is for research use only. |
| Batch-to-Batch Consistency | Filter-sterilized under controlled conditions; density controlled within 1.0770–1.0800 g/mL at 20 °C. |
For research use only, not for clinical use.
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