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| Product Name | Quick Exosome Isolation Kit Plus (for Milk) |
| Catalog No. | RESK-0614 |
| Description | Exosomes are small vesicles approximately 30-150 nm in diameter secreted by living cells, with a typical lipid bilayer structure. They are present in cell culture supernatant, serum, plasma, saliva, urine, amniotic fluid, and other biological fluids. Exosomes carry important cargo such as proteins, lipids, DNA, and RNA; they not only play important roles in intercellular material and information transfer, but are also promising early diagnostic biomarkers for a variety of diseases. This kit uses a unique isolation technology with optimized components for exosome extraction from milk, enabling fast and efficient recovery of exosome particles. The isolated exosomes can be used for electron microscopy analysis, NTA particle size analysis, nucleic acid analysis, protein analysis, cell-based experiments, and animal experiments. |
| Intended Use | Rapid and efficient isolation of exosomes from milk samples for downstream research applications. For research use only. |
| Principle / Technology | Optimized multi-step precipitation and filtration workflow: lipids and part of the proteins are first removed by centrifugation, contaminating proteins are then precipitated and clarified with Solutions A and B, the whey is filtered through a 50 mL centrifugal filter column (0.45 μm pore size), the buffer system is re-equilibrated with Solution C, and exosomes are finally precipitated with Solution D, collected by centrifugation, and resuspended. |
| Detection Method | Isolated exosomes can be analyzed by electron microscopy, NTA particle size analysis, nucleic acid analysis, and protein analysis, and used in cell-based and animal experiments. |
| Sample Type | Raw milk samples such as cow milk, goat milk, horse milk, and camel milk. Not recommended for milk powder samples. |
| Performance Range / Specifications | Recommended milk volume per extraction: 25-50 mL. Reagent dosages per 20 mL of whey: 2 mL of Solution A, 2.5 mL of Solution B, 2.5 mL of Solution C (same dose as Solution B), and 5 mL of Solution D; scale proportionally for other volumes. The 50 mL centrifugal filter column has a pore size of 0.45 μm. |
| Specificity | Optimized for exosome extraction from raw milk (cow, goat, horse, camel); not recommended for milk powder samples. |
| Reaction Conditions / Protocol | 1. Sample pretreatment 1) For frozen samples, thaw in a 25 °C water bath and place on ice once completely melted; for fresh samples, keep on ice after collection. 2) Recommended milk volume per extraction: 25-50 mL. 3) Defatting: centrifuge at 10,000 x g for 30 min at 4 °C to remove lipids and part of the proteins (after centrifugation the sample separates into three layers: an upper lipid layer, a lower protein pellet, and a middle whey layer; if the upper layer is soft and easily dislodged with a large pellet, repeat this step, collecting the middle layer each time). 4) Transfer the defatted whey (middle layer) to a new 50 mL centrifuge tube (a small amount of lipid and sediment carried over is normal and does not affect subsequent steps). 2. Removal of contaminating proteins 1) Whey clarification: add Solution A to the whey and mix by inversion, then add Solution B, mix by inversion, and let stand at 2-8 °C for 5 min (a bean curd-like solid with a transparent liquid phase should form; if not, add a little more Solution B). Per 20 mL of whey: 2 mL of Solution A and 2.5 mL of Solution B; scale proportionally. 2) Centrifuge the clarified whey at 10,000 x g for 20 min at 4 °C and collect the supernatant (if white particles are visible, repeat this step to avoid clogging in the next filtration). 3) Transfer the supernatant to a 50 mL centrifugal filter column and centrifuge at 3,000 x g for 10 min at 4 °C (repeat centrifugation until all liquid has passed through). 4) Buffer equilibration: transfer the filtrate to a new 50 mL tube, add Solution C (same dose as Solution B) and mix by inversion. 3. Exosome extraction 1) Add Solution D to the supernatant: 5 mL per 20 mL of whey; scale proportionally. 2) Cap the tube, mix by vortexing for 1 min, then let stand at 4 °C for at least 4 h (longer standing improves yield, but do not exceed 24 h). 3) Centrifuge at 10,000 x g for 30 min at 4 °C and discard the supernatant; the pellet is enriched in exosome particles. 4) Centrifuge again at 3,000 x g for 5 min at 4 °C and discard the residual supernatant. 5) Resuspend the pellet in an appropriate volume of 1×PBS (approximately 200 uL per 20 mL of whey) and transfer to a new 1.5 mL tube. 6) Centrifuge at 10,000 x g for 2 min at 4 °C and keep the supernatant, which is enriched in exosome particles (the exosome solution may appear slightly milky, which is normal). Note: for sterile processing, filter the exosomes through a 0.45 μm filter before use. |
| Components / Formulation | Solution A: 50 mL (store at 2-8 °C) Solution B: 70 mL (store at 2-8 °C) Solution C: 70 mL (store at 2-8 °C) Solution D: 110 mL (store at 2-8 °C) 50 mL centrifugal filter columns: 20 pcs (store at room temperature) |
| Storage Conditions | Store at 2-8 °C, placed upright. |
| Shelf Life | 1 year when stored as recommended. |
| Package Specifications | 20 T |
| Product Form | Liquid reagents with 50 mL centrifugal filter columns (0.45 μm pore size). |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Valid for 1 year when stored upright at 2-8 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Isolated exosomes are suitable for electron microscopy analysis, NTA particle size analysis, nucleic acid analysis, protein analysis, cell-based experiments, and animal experiments. |
| Recommended Buffer System | 1×PBS is recommended for resuspending the exosome pellet. |
| Application Notes / Precautions | Suitable for raw cow, goat, horse, and camel milk; not recommended for milk powder samples. After defatting centrifugation, the upper lipid layer should be compact and stable; if it is soft and easily dislodged, repeat the defatting step. If a sterile exosome preparation is required, filter through a 0.45 μm filter before use. The final exosome solution may appear slightly milky; this is normal. |
For research use only, not for clinical use.
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