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| Product Name | Serum Exosome Purification Kit, Precipitation Method, Ultracentrifugation-Free |
| Catalog No. | ATR-SPS-0029 |
| Description | Polymer-based precipitation kit for rapid isolation of intact exosomes and extracellular vesicles (EVs) from serum, plasma, cell culture conditioned medium, and urine without the need for ultracentrifugation. The proprietary precipitation reagent (PEG-based polymer solution) selectively reduces the solubility of exosomes and small EVs (30-200 nm diameter), enabling their pelleting by standard bench-top centrifugation (1,500 × g for 30 minutes). The gentle precipitation method preserves exosome surface protein integrity, making it suitable for downstream analyses including nanoparticle tracking analysis (NTA), electron microscopy, western blotting for exosomal markers (CD9, CD63, CD81, Alix, TSG101), RNA extraction, and functional studies. |
| Intended Use | Rapid and convenient isolation of intact exosomes and extracellular vesicles from small volumes (100 µL-1 mL) of biofluids and conditioned media for biomarker discovery, liquid biopsy research, drug delivery studies, and intercellular communication research. |
| Principle / Technology | PEG-based polymer reduces exosome solubility by volume exclusion effect; exosomes precipitate as a pellet at low centrifugal force (1,500 × g); gentle pelleting preserves vesicle membrane integrity; exosome pellet resuspended in PBS for downstream analysis. |
| Detection Method | Clarify sample by centrifugation (2,000 × g, 20 min) and/or 0.22 µm filtration; add Precipitation Reagent (1:5 ratio); mix by inversion; incubate at 2-8 °C for 1 hour to overnight; centrifuge 1,500 × g, 30 min, 4 °C; aspirate supernatant carefully; resuspend exosome pellet in PBS or lysis buffer. |
| Sample Type | Human and animal serum (100-500 µL), EDTA or citrate plasma, cell culture conditioned medium (1-10 mL, serum-free or exosome-depleted serum), urine (1-10 mL). |
| Performance Range / Specifications | Typical exosome yield: 50-500 µg protein from 1 mL serum; exosome size distribution 30-200 nm by NTA; particle concentration 10^9-10^11 particles/mL original sample. |
| Sensitivity / LOD | Exosomes detectable from as little as 100 µL serum; exosomal marker proteins (CD9, CD63, CD81) detectable by western blot from 10 µL serum equivalent. |
| Specificity | Preferentially precipitates particles of 30-200 nm diameter; predominantly exosomes/small EVs; some co-precipitation of larger microvesicles (>200 nm) and lipoprotein particles; higher purity than ultracentrifugation pellet with respect to exosomal protein markers. |
| Reaction Conditions / Protocol | Incubate sample with precipitation reagent at 2-8 °C; minimum 1 hour, recommended overnight for maximal yield; centrifuge at 1,500 × g; resuspend in buffer of choice. |
| Components / Formulation | Exosome Precipitation Reagent (sterile-filtered), sterile PBS (10 mL), detailed protocol with exosome characterization guidelines. |
| Storage Conditions | Store Precipitation Reagent at 2-8 °C; do not freeze. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 10 mL reagent (sufficient for ~50 serum/plasma samples at 200 µL each, or ~10 conditioned medium samples at 1 mL each), 50 mL bulk size. |
| Product Form | Clear, colorless to pale yellow viscous liquid. |
| Quality Control | Each lot tested for exosome precipitation efficiency from standardized human serum pool (protein yield >100 µg/mL serum), exosome marker enrichment (CD9/CD63 western >10-fold vs. serum), and NTA particle size distribution (mode within 50-150 nm). |
| Key Features | Ultracentrifugation-free protocol; standard bench-top centrifuge; 1-hour rapid precipitation; preserves exosome integrity; small sample volume (100 µL); PEG-based gentle method; scalable for high-throughput. |
| Purity | Precipitation reagent endotoxin <0.05 EU/mL; sterile-filtered 0.22 µm; DNase/RNase not detected. |
| Concentration | Ready-to-use; 200 µL reagent per 1 mL sample (1:5 ratio). |
| Activity / Unit Definition | Exosome precipitation efficiency >80% recovery of NTA particle count compared to 100,000 × g ultracentrifugation. |
| Molecular Weight | PEG polymer: average 6,000-8,000 Da. |
| Source / Origin | Synthetic PEG polymer in PBS buffer. |
| pH Range / Optimal pH | pH 7.2-7.4. |
| Shipping Conditions | Cold pack (2-8 °C); do not freeze. |
| Expiration Date / Stability | 12 months at 2-8 °C; after opening, use within 3 months under aseptic technique. |
| Regulatory / Compliance | For research use only; not for diagnostic procedures. |
| Compatibility | Compatible with serum, plasma (EDTA, citrate), cell culture conditioned medium (remove FBS exosomes by overnight ultracentrifugation or use exosome-depleted FBS), and urine. Not recommended for heparin plasma (heparin may interfere with precipitation). Lipemic samples may yield higher co-precipitated lipoprotein contamination. |
| Recommended Buffer System | PBS-based precipitation reagent, pH 7.3. |
| Application Notes / Precautions | Avoid hemolyzed serum/plasma samples — hemoglobin contamination may affect downstream assays. For conditioned medium, remove cells and debris by sequential centrifugation (300 × g 10 min, 2,000 × g 20 min) before adding reagent. Resuspend exosome pellet gently by pipetting (not vortexing). For RNA extraction, resuspend pellet directly in TRIzol. For functional assays, resuspend in sterile PBS and use fresh — do not freeze/thaw exosomes repeatedly as this damages vesicle integrity. Verify exosome isolation by western blot for at least two of: CD9, CD63, CD81, Alix, TSG101. |
| Batch-to-Batch Consistency | Precipitation efficiency within ±20% of reference lot based on total protein yield and CD63 western signal intensity. |
For research use only, not for clinical use.
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