Exosome Isolation and Purification Kit (Biofluids)

Cat. No.: E1526241

GRADE & PURITY BioReagent, Sterile

Overview

Exosomes are small vesicles (30–150 nm) secreted by cells, containing RNA and proteins, and are abundantly present in body fluids such as blood, saliva, urine and milk. Exosomes act as intercellular messengers to deliver effector molecules or signaling molecules between specific cells. Nevertheless, their structure, composition of effector substances and the biological pathways they participate in remain poorly characterized. Intact exosome particles are indispensable for research on the biological functions of exosomes. Conventional ultracentrifugation is plagued by complicated workflows, high requirements for specialized equipment and cumbersome operation. This exosome isolation and purification kit adopts optimized formulations, which is applicable to exosome extraction from body fluids including cerebrospinal fluid, amniotic fluid, milk, saliva, bronchoalveolar lavage fluid and nasal lavage fluid. Equipped with a purification filter device, the kit enables rapid and efficient acquisition of high-purity exosome pellets. The purified products can be applied to transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), nucleic acid analysis, protein analysis, cell-based assays and animal experiments.

Product Components

Component No.	Component Name	Appearance	3T	Storage
E1526241A	Exosome Concentration Solution	Liquid	1.8 mL	RT
E1526241B	Exosome Purification Filter	—	3 EA	RT

Required Materials

High-speed centrifuge, vortex mixer, 50 mL centrifuge tubes, 1.5 mL centrifuge tubes, sterile 1×PBS buffer.

Operating Protocol

I. Sample Pretreatment

1.Sample handling and thawing: For frozen samples, take them out from the refrigerator and thaw in a 25 °C water bath, then place fully thawed samples on ice. Fresh samples shall be placed on ice immediately after collection.

2.Recommended initial sample volume: For single isolation, the minimum recommended volume for bronchoalveolar lavage fluid, ascites and other body fluids is 1 mL; 2–3 mL is preferred per isolation run.

3.Centrifugation to remove cell debris: Transfer samples into centrifuge tubes and centrifuge at 3,000 ×g (~6,200 rpm*) for 10 min at 4 °C to remove cell debris. Note: If abundant pellets are observed, repeat centrifugation at 3,000 ×g for 10 min until no obvious precipitate exists, and retain the supernatant after each round of centrifugation.

*Calculated for small centrifuges with an effective radius of approximately 7 cm (for tubes ≤2 mL), the same applies hereinafter.

4.Centrifugation to remove impurity fragments: Transfer the supernatant to a new centrifuge tube and centrifuge at 12,000 ×g (~12,400 rpm) for 10 min at 4 °C to eliminate residual impurities and debris.

5.Supernatant transfer: Transfer the clarified impurity-free supernatant into a new centrifuge tube.

II. Exosome Isolation

1.Supernatant pretreatment: Add Exosome Concentration Solution (ECS reagent) to the clarified supernatant. Refer to the table below for detailed dosing standards; volumes for other sample input amounts can be calculated proportionally based on the listed ratios.

Sample Name	Sample Volume	ECS Volume
Bronchoalveolar Lavage Fluid (BALF)	1 mL	0.25 mL
Ascites	1 mL	0.25 mL

2. Solution Mixing: After adding ECS reagent, tightly cap the centrifuge tube and vortex thoroughly for 1 minute on a vortex mixer. Incubate the mixture at 4 °C for no less than 8 hours. Note: Extended incubation time can improve exosome yield, but the incubation duration must not exceed 24 hours.

3. Exosome Precipitation: Take out the tube containing the mixture and centrifuge at 12,000 ×g (~12,400 rpm) for 30 minutes at 4 °C. Discard the supernatant completely to the greatest extent possible; the pellet is enriched with exosome particles.

4. Secondary Short Spin: Centrifuge the tube with the pellet again at 12,000 ×g (~12,400 rpm) for 2 minutes at 4 °C, then thoroughly remove all residual supernatant.

5. Exosome Resuspension: Gently pipette the pellet up and down with an appropriate volume of 1×PBS buffer until fully dissolved, then transfer the resuspension to a new 1.5 mL centrifuge tube. Recommended resuspension volume: approximately 100 μL of 1×PBS for every 1 mL starting body fluid sample.

6. Harvest Crude Exosome Pellets: Centrifuge the 1.5 mL tube with resuspended sample at 12,000 ×g (~12,400 rpm) for 2 minutes at 4 °C, retain the supernatant which contains enriched crude exosomes. Note: If visible precipitate remains, repeat centrifugation at 12,000 ×g for 2 minutes until no obvious pellet is observed, and collect the supernatant after each centrifugation.

III. Exosome Purification

1. Column Purification: Combine all crude exosome suspensions derived from the same type of body fluid and load them into the upper chamber of the Exosome Purification Filter (EPF column). Centrifuge at 3,000 ×g (~6,200 rpm) for 10 minutes at 4 °C. Collect the eluate at the bottom of the EPF column, which is the final purified exosome product. Note: EPF columns are disposable and cannot be reused.

2. Exosome Storage: Aliquot the purified exosomes into appropriate volumes and store long-term in a -80 °C freezer for subsequent downstream experiments.

Note:

This product is for life science research use only. It shall not be applied for medical diagnosis or any clinical purposes.

Specifications

Synonyms

Body Fluid-Derived Exosome Extraction & Purification Kit

Specifications & Purity

BioReagent, Sterile

Grade

BioReagent, Sterile

Application

Sample type

body fluids

Storage and Shipping

Storage

Room temperature

Shipped In

Normal

Stability And Storage

Store at RT long term (24 months).

Contents & Storage

Component No.	Component Name	Appearance	3T	Storage
E1526241A	Exosome Concentration Solution	Liquid	1.8 mL	RT
E1526241B	Exosome Purification Filter	—	3 EA	RT

Application Protocols

Item-specific tested applications, recommended dilutions/inputs, and positive controls: Not specified for this item; refer to CoA/Spec Sheet and the product’s Instructions for Use.

Note: While general exosome isolation workflows typically include sample pre-clearing, incubation with isolation reagent, capture/recovery by centrifugation or affinity, washing, and elution/resuspension for downstream assays, those steps and parameters must be confirmed against the official instructions for this kit.

Biological Roles

General background (literature; for context only)

• Exosomes are nanoscale extracellular vesicles (~30–150 nm) released by most cell types via the endosomal (multivesicular body) pathway. They carry selective cargo—proteins (e.g., tetraspanins CD9/CD63/CD81), lipids (ceramides, cholesterol), and nucleic acids (miRNA, mRNA fragments, DNA)—reflecting cell state and origin.
• Functions in intercellular communication:
  • Transfer of functional proteins and RNAs influencing recipient cell signaling and gene expression.
  • Participation in immune modulation, development, and tissue homeostasis; their content and abundance change with physiological and environmental cues.
• Biochemical signatures:
  • Enrichment of late endosome markers (Alix/PDCD6IP, TSG101), ESCRT components, specific lipids (sphingomyelin-rich rafts), and post-translational modifications.
  • Density and size distinguish exosomes from other EVs (microvesicles, apoptotic bodies), though overlap exists; rigorous characterization per MISEV guidelines is recommended.

Research implications

• Isolated exosomes enable discovery research in cell-to-cell communication, biomarker exploration, and cargo functional studies. Purity and integrity influence interpretability of downstream omics measurements; thus, isolation kits are paired with orthogonal characterization (NTA, EM, Western blot) to confirm EV identity and assess co-isolates such as lipoproteins or protein aggregates.

Item-specific note

• This kit supports exosome isolation from biofluids for research use only; no clinical diagnostic use is implied.

Buffer Applications

Item-specific

• Buffer identities, compositions, and pH ranges supplied in this kit: Not specified for this item; refer to CoA/Spec Sheet and component labels.

General guidance for exosome workflows (literature)

• Pre-clearing and dilution: Use sterile, nuclease-free PBS or isotonic buffers (pH ~7.2–7.4) to dilute viscous biofluids (e.g., serum) and reduce matrix effects prior to isolation.
• Washing: Isotonic, calcium/magnesium-free buffers can reduce aggregation during wash steps. Gentle mixing preserves vesicle integrity.
• RNA-compatible handling: Use RNase-free buffers and consumables; avoid DEPC with amine-containing components. For RNA extraction from isolated EVs, employ chaotropic lysis buffers immediately after capture to stabilize nucleic acids.
• SEC polishing (optional): If purity is critical, post-isolation SEC with phosphate or HEPES-buffered saline (pH 7.2–7.4) separates EVs from polymers and protein co-isolates; collect fractions by UV or particle counts.
• Storage buffer: For short-term storage of purified EVs, sterile PBS with cryoprotectants (e.g., trehalose, literature practice) can mitigate aggregation; validate compatibility with downstream assays. Note: This is general guidance, not a specification for this kit.

Green Alternatives

General sustainability considerations (literature)

• Method choice vs. environmental impact:
  • Precipitation chemistry typically avoids energy-intensive ultracentrifugation, reducing instrument energy use and lab time. Trade-off: potential co-precipitation increases need for additional cleanup steps.
  • Size-exclusion chromatography (SEC) uses aqueous buffers with minimal hazardous waste; produces higher purity at the cost of consumables and buffer volumes.
  • Affinity capture concentrates EVs rapidly with low energy input; resin/bead reuse policies (if compatible) can reduce waste.

Comparison (general; not item-specific)

• Precipitation kits: low energy, minimal hardware; generate polymer-containing aqueous waste—verify local disposal rules.
• Ultracentrifugation: high energy and capital cost; low chemical waste but larger time footprint.
• Microfluidic/filtration approaches: low reagent volumes; plastic consumable waste can be significant.

Greener practices

• Use reusable glassware where sterility permits; minimize single-use plastics while maintaining RNase/DNase-free conditions.
• Optimize batch sizing to reduce leftover reagents; prepare only the volumes needed for the day’s work.
• Buffer preparation with concentrated stocks minimizes shipping weight and packaging. Validate that concentration/dilution does not alter isolation performance.
• Plan orthogonal QC (NTA/SEC) to avoid repeating isolations and excess reagent consumption.

Pharmaceutical Uses

Not applicable to this research-use-only kit. No pharmacopeial status or GMP manufacturing claims are made. In a general R&D context (literature), exosome preparations may be evaluated as materials for analytical method development or as model vesicles; however, this product is not intended for clinical or therapeutic applications, patient diagnostics, or as a drug substance/excipient.

Physical Properties

Item-specific (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula / molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Other specifications (pH of buffers, refractive index, density): Not specified for this item; refer to CoA/Spec Sheet.

General/literature context for exosome kits (not product specifications)

• Kit reagents are commonly provided as aqueous buffers and/or polymer/affinity solutions; viscosity and pH are method-dependent (e.g., polymer precipitation solutions are often viscous; affinity buffers are typically near physiological pH).
• Storage stability at room temperature can indicate inclusion of preservatives or lyophilized formats; exact details for this item are Not specified for this item.
• Exosome physical attributes (for downstream QC, literature values):
  • Typical hydrodynamic diameter: ~30–150 nm (NTA/DLS)
  • Buoyant density (sucrose/D2O gradients): ~1.10–1.19 g/mL
  • Zeta potential: often negative (−10 to −30 mV), buffer dependent These are general expectations for exosome preparations and not specifications of this kit.

Quality & Grades

Item-specific (from Product Data)

• Grade/Purity: BioReagent, Sterile.
  • BioReagent: Indicates suitability for laboratory research workflows. Not intended for clinical diagnostics or GMP manufacturing use.
  • Sterile: Components are supplied sterile to minimize microbial contamination of preparations, important for preserving vesicle integrity and preventing nuclease/protease contamination. Sterility does not imply apyrogenicity; endotoxin content is Not specified for this item; refer to CoA/Spec Sheet.
• Research use note: For research use only.

General guidance (literature)

• Quality attributes relevant to exosome isolation kits include: batch-to-batch consistency, nuclease/protease-free status, particulate/bioburden control, and compatibility with common downstream assays (NTA, TEM/cry-EM, Western blot for tetraspanins, LC–MS proteomics, RNA-seq). Specific performance metrics for this item are Not specified for this item; refer to CoA/Spec Sheet.
• Documentation: For method validation, obtain the lot-specific Certificate of Analysis (CoA) for any available acceptance criteria (e.g., sterility test, functional recovery benchmarks) and component expiry dates.
• Practical considerations:
  • If analyzing RNA cargo, confirm RNase-free handling and consider using RNase inhibitors post-isolation.
  • For proteomics, use MS-compatible buffers or perform buffer exchange post-isolation to remove polymers/surfactants that can suppress ionization.

Reaction & Applications

Item-specific (from Product Data)

• Intended use: Exosome isolation and purification from biofluids for research use only.

General applications (literature; not product-validated claims)

• Source matrices: human/animal serum or plasma, urine, CSF, saliva, and conditioned cell culture media. Pre-clearing (low-speed spins, 0.22–0.45 μm filtration) reduces cells/debris before isolation.
• Downstream analyses of isolated exosomes:
  • Particle characterization: nanoparticle tracking analysis (NTA), tunable resistive pulse sensing (TRPS), DLS.
  • Morphology: TEM/cry-EM, AFM.
  • Protein profiling: Western blot (CD9/CD63/CD81, Alix, TSG101), ELISA, flow cytometry (bead-coupled EVs), LC–MS-based proteomics.
  • Nucleic acids: miRNA/lncRNA profiling by RT-qPCR or RNA-seq; DNA analyses as applicable.
  • Lipidomics: LC–MS of EV lipidome after solvent extraction.
• Methodological considerations:
  • Precipitation-based kits: simple and scalable; may co-precipitate protein/lipoproteins—include appropriate controls and optional clean-up (size-exclusion chromatography, SEC) for purity-critical assays.
  • Affinity-based capture (e.g., tetraspanin-targeting resins/beads): higher specificity; may bias toward subpopulations.
  • Differential ultracentrifugation/SEC (complementary methods): used for orthogonal validation of purity and size distribution.
• Practical tips (general): keep samples cold during pre-clearing to preserve vesicle integrity; avoid repeated freeze–thaw of biofluids; document input volume, recovery (particles/mL), and protein-to-particle ratios to assess preparation quality.

Reaction Conditions

Not applicable as a chemical reaction reagent. For method planning (general, literature-based):

• Temperature: Perform biofluid handling on ice or at 4°C during pre-clearing to preserve vesicles; follow kit-specific instructions for incubation temperatures/times (Not specified for this item).
• Incubation: Precipitation chemistries often require 30–120 min at low temperature; affinity captures may range 15–120 min with gentle mixing. Exact conditions for this item are Not specified for this item; refer to CoA/Spec Sheet or Instructions for Use.
• Centrifugation/filtration: Low-to-moderate g-forces (e.g., 1,500–10,000 × g) are common for clearing and pelleting in kit-based methods; verify rotor limits and tube compatibility.
• Yield/purity: Highly matrix- and method-dependent; benchmark using orthogonal QC (particle counts, protein assays, immunomarkers). These are general guidelines and not performance claims for this kit.

Safety & Handling

Item-specific (from Product Data)

• GHS classification, H-statements, pictograms, signal word: Not specified for this item; refer to SDS for each component.
• Storage conditions: Room temperature (as supplied). Protect from contamination. Avoid extreme temperatures unless otherwise noted on component labels.

General safety guidance (literature; defer to SDS)

• Personal protective equipment: lab coat, safety glasses, and appropriate gloves (e.g., nitrile). Use in a clean area to avoid environmental particulate contamination of vesicle preps.
• Biofluid handling: Treat all human- or animal-derived biofluids as potentially infectious. Follow BSL-2 practices where applicable and institutional biosafety rules. Use splash protection and work in a certified biological safety cabinet when aerosol generation is possible.
• Chemical hazards: Precipitation or affinity reagents may contain salts, polymers, or preservatives. Avoid inhalation/ingestion; prevent contact with skin and eyes. Consult SDS for incompatibilities (e.g., strong oxidizers, acids/bases) and spill response.
• Special risks and tips:
  • Avoid microbial contamination; sterile, RNase/DNase-free technique is recommended, particularly for RNA cargo analysis.
  • Temperature swings can cause salt crystallization or phase separation in some reagents; bring to room temperature and mix gently to re-dissolve before use.
  • Waste disposal: Collect biohazardous waste (spent biofluids, disposables) per institutional and regulatory requirements; segregate chemical waste from biological waste.
• First-aid overview: In case of skin/eye contact, rinse with water for at least 15 minutes; seek medical attention if irritation persists. If inhaled or ingested, seek medical advice. Refer to SDS.

Solvent Selection

Not typically applicable. This product is a multi-component isolation kit rather than a solvent. If downstream processing requires solvent-based steps (e.g., lipid extraction), select solvents according to the analytical method (e.g., chloroform/methanol for Bligh–Dyer or MTBE-based extractions) and ensure compatibility with the isolation chemistry. For general aqueous handling, use nuclease-free, sterile water and buffers.

Storage & Reconstitution

Item-specific (from Product Data)

• Storage conditions (as supplied): Room temperature. Protect from contamination and moisture. Follow any component-specific labels that may differ.
• Shipping: Normal.
• Reconstitution details (e.g., for lyophilized components) and working solution stability: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature)

• Before first use: Inspect components for precipitation or crystallization; if observed, equilibrate to room temperature and mix gently to re-dissolve. Do not heat above recommended temperatures.
• Aseptic handling: Maintain sterility when opening containers; use sterile, nuclease-free pipette tips and tubes.
• Working aliquots: If components are compatible, prepare single-use aliquots to minimize contamination. Record open dates and adhere to component expiry.
• Storage of isolated exosomes (post-purification, general practice):
  • Short term (≤1 week): 2–8°C, minimize headspace; avoid repeated freeze–thaw.
  • Longer term: −80°C with cryoprotectant (e.g., trehalose, literature). Validate stability for intended assays. These are general recommendations only; defer to the kit’s official Instructions for Use and CoA for definitive guidance.

Structure & Identity

Item-specific (from Product Data)

• SKU: E1526241
• Product name: Exosome Isolation and Purification Kit (Biofluids)
• Category: Life Science research reagent kit
• Grade/Purity: BioReagent, Sterile
• Storage: Room temperature (as supplied)
• Shipping: Normal
• Chemical identifiers (CAS, CID, InChIKey, SMILES, molecular formula, molecular weight): Not specified for this item; refer to CoA/Spec Sheet.

General description (literature)

• This product is a multi-component kit intended to isolate and purify extracellular vesicles (exosomes) from biofluids (e.g., serum, plasma, urine, cerebrospinal fluid, saliva) for downstream research analyses.
• Typical kit components in the market include selective precipitation or affinity capture reagents, wash/lysis buffers, and accessory consumables; the exact composition for this item is Not specified for this item; refer to CoA/Spec Sheet.
• Structural features: Not applicable—this is a kit, not a single small molecule. No single 2D/3D chemical structure applies.

2D structure (explanatory)

• Not applicable. Exosomes are nanoscale lipid bilayer vesicles (~30–150 nm), heterogeneous in surface proteins and cargo; a single structural formula cannot represent the kit contents.

Synthetic Utility

Not typically applicable. This product is a biological isolation kit rather than a synthetic reagent or building block. It does not participate in organic transformations or polymerizations. For chemists integrating EV studies with synthesis (e.g., lipid labeling or click-chemistry tagging of vesicles), perform such reactions on isolated materials using orthogonal, biocompatible chemistries and validate that residual kit reagents do not interfere with coupling efficiency or analyses.

Target Specificity

No antibody or probe specificity is defined for this product. Target antigens, epitopes, species reactivity, clone, and isotype: Not applicable/not specified for this item.