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| Product Name | Quick Exosome Isolation Kit Plus (for Cell Culture Media) |
| Catalog No. | RESK-0610 |
| 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 to rapidly obtain large amounts of intact exosome particles from cell culture supernatant. The isolated exosomes are suitable for downstream applications such as cell co-culture, electron microscopy analysis, Western blot, quantitative PCR (qPCR), and high-throughput sequencing. |
| Intended Use | Rapid isolation and purification of intact exosomes from cell culture supernatant for downstream research applications. For research use only. |
| Principle / Technology | Precipitation-based exosome enrichment combined with column purification: an exosome extraction reagent is mixed with the pre-cleared supernatant and incubated at 4 °C to precipitate exosomes, which are collected by centrifugation, resuspended, and further purified on an EPF purification column (a 220 nm filter column that removes large-particle impurities). |
| Detection Method | Isolated exosomes can be analyzed by electron microscopy, nanoparticle tracking analysis, Western blot, qPCR, and high-throughput sequencing. |
| Sample Type | Cell culture supernatant (this kit is intended only for cell culture supernatant; it is not suitable for exosome extraction from serum/plasma). |
| Performance Range / Specifications | Recommended initial sample volume per extraction: not less than 20 mL of cell culture supernatant. Reagent-to-sample ratio: 1:4; with a 50 mL centrifuge tube, the maximum sample load is 40 mL. One reaction (1 T) processes 20-30 mL of cell culture supernatant; for tumor cells, no less than 20 mL is recommended. For stem cell supernatant, concentrate 3-5 fold by ultrafiltration before extraction (i.e., 60-100 mL of supernatant). Yield per 1 mL of sample: approximately 1-2 μg of protein and 0.1-0.4 ng of RNA. |
| Specificity | Designed specifically for exosome isolation from cell culture supernatant; not applicable to serum/plasma samples. |
| Reaction Conditions / Protocol | 1. Sample pretreatment 1) Collect fresh samples and keep on ice; for frozen samples, thaw in a 25 °C water bath and place on ice once completely melted. 2) Recommended initial sample volume per extraction: not less than 20 mL of cell culture supernatant. 3) Centrifuge at 3,000 x g for 10 min at 4 °C to remove cell debris (if the pellet is large, repeat 3,000 x g for 10 min until no obvious pellet remains, keeping the supernatant each time). 4) Transfer the supernatant to a new tube and centrifuge at 10,000 x g for 10 min at 4 °C to remove impurity debris. 5) Transfer the cleared supernatant to a new centrifuge tube. 2. Exosome extraction 1) Add the exosome extraction reagent (Reagent A) to the cleared supernatant at a reagent-to-sample ratio of 1:4; scale proportionally for other sample volumes. 2) Cap the tube, mix by vortexing for 1 min, then let stand at 4 °C for at least 8 h (longer standing improves yield, but do not exceed 24 h). 3) Centrifuge at 10,000 x g for 60 min at 4 °C and discard the supernatant (aspirate as completely as possible). 4) Centrifuge again at 10,000 x g for 2 min at 4 °C and discard the residual supernatant. 5) Resuspend the pellet in an appropriate volume of 1×PBS by gentle pipetting (approximately 200 uL of 1×PBS per 20 mL of cell culture supernatant) and transfer to a new 1.5 mL tube. 6) Centrifuge at 12,000 x g for 2 min at 4 °C and keep the supernatant, which is enriched in exosome particles (if the pellet is large, repeat until no obvious pellet remains). 3. Exosome purification 1) Transfer the harvested crude exosome preparation into the upper chamber of the EPF purification column and centrifuge at 3,000 x g for 10 min at 4 °C; collect the liquid at the bottom of the column tube, which contains the purified exosome particles (the EPF column is single-use and cannot be reused). 2) Aliquot the purified exosomes and store at -80 °C for subsequent experiments. |
| Components / Formulation | Exosome extraction reagent (Reagent A) and EPF purification columns (220 nm filter columns). |
| Storage Conditions | Store at room temperature. |
| Shelf Life | 2 years when stored as recommended. |
| Package Specifications | 10 T 20 T 2 T |
| Product Form | Liquid reagent with single-use purification columns. |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Valid for 2 years at room temperature. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Isolated exosomes are suitable for cell co-culture, electron microscopy, Western blot, qPCR, and high-throughput sequencing. Can be combined with an exosome ultrafiltration tube to further improve purity. |
| Recommended Buffer System | 1×PBS is recommended for resuspending the exosome pellet. |
| Application Notes / Precautions | To ensure that the obtained exosomes originate from your cells, it is best to culture cells with exosome-depleted serum. For further purification, antibody-coated magnetic beads can be used for affinity purification. This kit is intended only for exosome isolation from cell culture supernatant and is not suitable for serum/plasma; please use the serum/plasma exosome isolation kit for those samples. The purity of precipitation-based isolation is relatively low; combining with an exosome ultrafiltration tube is recommended. The precipitation method does not affect previously labeled (stained) exosomes, and labeled exosomes can be re-extracted with reagent added proportionally to the sample volume; there is no upper limit on the amount of exosomes re-extracted. For your safety and health, wear a lab coat and disposable gloves during operation. |
For research use only, not for clinical use.
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