Human peripheral blood lymphocyte separation solution
Clearly layered, sterile and low-toxicity.
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Instruction manual- Product Description
- Composition
- Precautions
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[Product Introduction]
Human lymphocyte separation solution is ready-to-use and designed for isolating lymphocytes from human peripheral blood. This solution contains polysucrose and diatrizoate, which enhance the efficiency of lymphocyte isolation. The product is supplied in sterile packaging and has been tested for endotoxins; the isolated cells, after washing, are suitable for in vitro culture without compromising subsequent experimental procedures. The underlying principle relies on the differences in cell size, morphology, density, and specific gravity among the various cell types in human peripheral blood to achieve selective separation.
[Product Information]
Specification: 250 mL/bottle
Endotoxin content < 0.5 EU/mL
Separate liquid density: (1.077 ± 0.001) g/mL (20°C); Osmolality: (290 ± 15) mOsmol/kg.
Storage conditions: Store at 4°C–30°C, protected from light; shelf life: 3 years. Transport at room temperature.
[Instructions for Use]
1. Collect fresh anticoagulated whole blood (using EDTA, sodium citrate, or heparin as the anticoagulant). Dilute the whole blood with an equal volume of an isotonic solution (PBS or saline).
2. Add an appropriate volume of separation solution to the centrifuge tube (the total volume should not exceed two-thirds of the tube’s capacity, as this may compromise the separation efficiency). Carefully layer the diluted blood sample on top of the separation solution, ensuring a clear interface between the two phases. The ratio of separation solution, undiluted anticoagulated whole blood, and an isotonic solution (PBS or saline) should be 1:1:1.
3. At room temperature, centrifuge with a horizontal rotor at 700–800 g for 20–30 minutes.
4. After centrifugation, the bottom of the tube contains red blood cells, the middle layer consists of the separation medium, and the top layer is the plasma/tissue homogenate. Between the plasma layer and the separation medium lies a thin, relatively dense white layer—namely, the mononuclear cell layer, which includes lymphocytes and monocytes. Carefully aspirate the white layer into another centrifuge tube.
5. Dilute to a specified volume with an isotonic solution (PBS, saline, or culture medium), and invert to mix thoroughly. At room temperature, centrifuge in a horizontal rotor at 250g for 10 minutes, then discard the supernatant. Repeat the washing step 1–2 times.
6. Resuspend the cells in an isotonic solution (PBS, saline, or culture medium) for later use.
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[Precautions]
1. This product is light-sensitive and should be stored at room temperature, protected from light.
2. Please allow the lymphocyte separation solution to equilibrate to room temperature (18°C–30°C) prior to use. Before opening the vial, invert it several times to mix thoroughly. Perform all procedures under aseptic conditions to prevent contamination.
3. To maintain lymphocyte viability, blood samples should ideally be fresh peripheral blood from healthy individuals.
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Goonie Biotechnology (Guangzhou) Co., Ltd.
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