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| Product Name | Lymphocyte Separation Medium 1.084 |
| Catalog No. | RESK-0257 |
| Description | Lymphocyte Separation Medium 1.084 (LSM-1.084) is a sterile, ready-to-use separation medium, an improved formulation based on the classic Boyum polysucrose-sodium diatrizoate method that makes the procedure simple and rapid with higher separation efficiency. It is a sterile mixed solution of polysucrose and sodium diatrizoate with a density of 1.084 g/mL and a very low endotoxin level ( |
| Intended Use | Isolation of higher-density human mononuclear cells (lymphocytes and monocytes) from peripheral blood, umbilical cord blood and bone marrow-derived samples Isolation of mouse and rat lymphocytes/blood cells (rodent lymphocytes are slightly denser than human lymphocytes) Isolation of granulocytes |
| Principle / Technology | Density gradient centrifugation. Diluted blood is layered onto the 1.084 g/mL medium; during centrifugation, erythrocytes and polymorphonuclear leukocytes are pelleted while mononuclear cells (lymphocytes and monocytes) collect as a distinct band at the plasma-medium interface. |
| Detection Method | Density gradient centrifugation at 400 × g for 30-40 min at room temperature, followed by collection of the mononuclear cell band and washing steps. |
| Sample Type | Defibrinated or anticoagulated human peripheral blood, umbilical cord blood and bone marrow-derived tissue samples; mouse and rat blood. Heparin, EDTA, sodium citrate, ACD or CPD anticoagulants are all acceptable; defibrinated blood requires no anticoagulant. |
| Performance Range / Specifications | Density: 1.084 g/mL Endotoxin: <0.12 EU/mL Sterile, ready-to-use mixed solution of polysucrose and sodium diatrizoate Manufactured under a certified quality management system |
| Specificity | Optimized density of 1.084 g/mL for human mononuclear cells denser than those recovered with 1.077 g/mL media, and for rodent (mouse/rat) lymphocytes. |
| Reaction Conditions / Protocol | 1. Equilibrate LSM-1.084 at room temperature for 10-30 min before use. 2. Dilute 2 mL of defibrinated or anticoagulated blood with an equal volume (2 mL) of sterile balanced salt solution (1:1); mix by inverting in a 10-15 mL centrifuge tube or by gentle pipetting. Larger blood volumes give the same separation efficiency if wider tubes are used to maintain a blood layer height of about 3 cm over about 2.4 cm of LSM-1.084. 3. Gently invert the bottle to mix LSM-1.084 thoroughly; aseptically transfer 3 mL into a 10-15 mL centrifuge tube using a syringe or pipette tip. 4. Carefully layer the diluted blood on top of the LSM-1.084 (room temperature) to form a clear interface; do not mix the blood into the medium. 5. Centrifuge at 400 × g for 30-40 min at room temperature. Erythrocytes and polymorphonuclear leukocytes are pelleted, and a mononuclear cell band forms at the interface (from top to bottom: plasma - mononuclear cell layer - LSM-1.084 layer - granulocytes - erythrocytes). 6. With a sterile tip, aspirate the platelet/plasma layer 2-3 mm above the mononuclear cell band, taking care not to disturb the band (alternatively, collect the band directly; the upper cell-free plasma may be retained for other uses). 7. Transfer the entire mononuclear cell layer, together with a small amount of overlying plasma and underlying LSM-1.084, into a new sterile centrifuge tube. 8. Add at least 3 volumes (about 6 mL) of balanced salt solution and resuspend the cells thoroughly by gentle pipetting. 9. Centrifuge at 400-500 × g for 10-15 min at room temperature. Higher speed improves recovery; if thorough platelet removal is required, use low-speed centrifugation (60-100 × g). 10. Discard the supernatant and resuspend the mononuclear cells in 6-8 mL of balanced salt solution. 11. Centrifuge at 400-500 × g (or 60-100 × g for platelet removal) for 10 min at room temperature. 12. Discard the supernatant and resuspend the mononuclear cells in an appropriate medium for use. Granulocyte isolation: follow steps 1-6 above; carefully aspirate the LSM-1.084 layer without drawing in the granulocyte layer; transfer the white granulocyte layer (above the erythrocyte layer) into a sterile 50 mL centrifuge tube; resuspend in at least 5 volumes of balanced salt solution and centrifuge at 400 × g for 15 min; discard the supernatant, wash once more in 6-8 mL of balanced salt solution at 400-500 × g for 10 min at room temperature; discard the supernatant and resuspend the granulocytes in an appropriate medium. |
| Components / Formulation | Sterile aqueous mixture of polysucrose and sodium diatrizoate, density 1.084 g/mL; ready-to-use. |
| Storage Conditions | Store sealed and protected from light at 4-30 °C. After opening, store at 4-8 °C. |
| Shelf Life | 3 years when stored sealed at 4-30 °C, protected from light. |
| Package Specifications | 100 mL |
| Product Form | Sterile, ready-to-use liquid |
| Quality Control | Sterility tested; endotoxin <0.12 EU/mL; density controlled at 1.084 g/mL; produced in a strictly controlled environment under a certified quality management system. |
| Shipping Conditions | Room temperature. |
| Expiration Date / Stability | Stable for 3 years when stored sealed and protected from light at 4-30 °C; after opening, keep at 4-8 °C. |
| Regulatory / Compliance | Manufactured under a certified quality management system. |
| Recommended Buffer System | Balanced salt solutions for dilution and washing: a commercial balanced salt solution is recommended; Ca2+/Mg2+-free phosphate buffer (e.g., DPBS), Hank's solution, or cell culture medium (e.g., RPMI 1640) can also be used. |
| Application Notes / Precautions | Equilibrate the medium to room temperature for 10-30 min before use and mix by gentle inversion. Layer the diluted blood carefully to maintain a sharp interface; do not mix it into the medium. Collect the entire mononuclear cell band together with a small amount of overlying plasma and underlying medium to maximize recovery. Use low-speed centrifugation (60-100 × g) during washing if complete platelet removal is required. |
For research use only, not for clinical use.
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