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| Product Name | Polysucrose 400 |
| Catalog No. | RESK-0254 |
| Description | Polysucrose 400 is a highly branched, completely non-ionic polymer formed by the copolymerization of sucrose and epichlorohydrin. Rich in hydroxyl groups, it is strongly hydrophilic and readily soluble in water. It is commonly used as a density gradient medium for the separation and isolation of eukaryotic cells, organelles and bacterial cells, and as a stabilizer and separation medium for lymphocyte separation, with densities up to 1.2 g/mL. In immunological research, it can also serve as a hapten carrier. In concentration dialysis, its high molecular weight prevents passage through dialysis membranes, making it effective for concentrating sensitive materials. In physiological and cell stability studies, it can be used to monitor protein excretion in blood vessels, and it can be added to electrophoresis solutions as an electrolyte stabilizer. Due to its high molecular weight and lower content of dialyzable substances compared with sucrose, polysucrose 400 generally does not penetrate cell membranes during cell separation, better preserving cell function and morphology. |
| Intended Use | Density gradient medium for separation and isolation of eukaryotic cells, organelles and bacterial cells; separation of cells sensitive to centrifugal pelleting and of cells with similar densities but different sizes; lymphocyte separation (as a component of lymphocyte separation media); hapten carrier in immunology; concentrating agent for sensitive materials in dialysis; electrolyte stabilizer in electrophoresis. For research use only. |
| Principle / Technology | A neutral, highly branched sucrose-epichlorohydrin copolymer that forms aqueous density gradients (up to 1.2 g/mL) for continuous or discontinuous gradient centrifugation and unit-gravity sedimentation. Particles band at positions matching their buoyant densities, enabling separation without penetrating cell membranes. |
| Sample Type | Eukaryotic cells, organelles, bacterial cells, lymphocytes |
| Performance Range / Specifications | Density range: up to 1.2 g/mL; most cells and organelles have buoyant densities of 1.0–1.2 g/mL in this medium. Non-ionic; strongly hydrophilic; low content of dialyzable substances. Representative buoyant densities: membranes 1.05 (100,000 × g, 16 h); pigment cells 1.07 (195,000 × g, 36 h); hepatocytes 1.10–1.15 (6,000 × g, 2 h); fibroblasts 1.05 (8,000 × g, 1 h); Ehrlich ascites carcinoma cells 1.07 (1,400 × g, 45 min). |
| Specificity | Non-ionic polymer; does not penetrate cell membranes; metabolically inert and non-cytotoxic. |
| Reaction Conditions / Protocol | Applicable to discontinuous and continuous gradient centrifugation as well as unit-gravity separation. Advantages of discontinuous gradient centrifugation: a) the abrupt density change between layers concentrates separated cells into sharp bands at the interfaces, allowing easy removal with a pipette; b) cells with very different densities can be rapidly separated using only two density layers. Preparation of discontinuous gradients: 1. Prepare solutions in a suitable buffer or isotonic 0.25 M sucrose solution at concentrations appropriate for the target material; most cells and organelles have buoyant densities between 1.0 and 1.2 g/mL, and two layers usually suffice. Prepared solutions can be stored in the refrigerator but should be used at room temperature. 2. Layer the gradient solutions into standard centrifuge tubes (about 1 cm deep per layer), with the densest layer at the bottom. 3. Carefully place the sample on the top layer and gently stir the interface with a glass rod to eliminate the boundary. Centrifuge; particles of different densities will collect within or between the layers. 4. After centrifugation, remove each phase, recover the target cells, and wash with a suitable buffer to remove the polymer. Repeat the wash 4–5 times. |
| Components / Formulation | Copolymer of sucrose and epichlorohydrin; highly branched, completely non-ionic; rich in hydroxyl groups; strongly hydrophilic and readily soluble in water. |
| Storage Conditions | Store at room temperature, protected from light. |
| Shelf Life | At least 5 years |
| Package Specifications | 25 g |
| Product Form | Powder |
| Purity | Low content of dialyzable substances compared with sucrose |
| Molecular Weight | Approximately 400,000 Da |
| Source / Origin | Synthetic copolymer of sucrose and epichlorohydrin |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | At least 5 years when stored at room temperature protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with continuous and discontinuous gradient centrifugation and unit-gravity sedimentation; suitable for cells sensitive to centrifugal pelleting. Residual polymer can be estimated by the anthrone reaction. |
| Recommended Buffer System | Suitable buffers or isotonic 0.25 M sucrose solution. |
| Application Notes / Precautions | Prepared gradient solutions can be stored in the refrigerator but should be used at room temperature. In some cases, continuous or linear density gradients may be required; for simple separations, a homogeneous solution without a gradient can be used with stepwise increases in centrifugation speed. Unit-gravity sedimentation is widely used to separate centrifugation-sensitive cells, and cells with similar densities but different sizes can also be efficiently separated by this method. Wash separated cells 4–5 times with suitable buffer to remove residual polymer; residual amounts can be estimated by the anthrone reaction. |
For research use only, not for clinical use.
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