Exosome isolation and analysis | Miltenyi Biotec

Application protocol for isolation and analysis of human exosomes

Protocol

Isolation and analysis of human exosomes

Exosome isolation

Exosomes belong the to group of extracellular vesicles (EV) and play a key role in cellular functions. Consequently, they are of great interest in basic research as well as in future diagnostic applications or clinical settings. We offer a reliable isolation method that safes time, capital equipment, and high starting volumes (50–100 mL) – a true alternative for conventional ultracentrifugation.

Principle of magnetic isolation of EVs using the Exosome Isolation Kit

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Isolation of EVs using MACS® Technology

Using our Exosome Isolation Kit, enrichment of EVs has never been easier.

Selection of EVs according to surface markers i.e. CD9, CD63, or CD81
Low starting volume of samples (0.5 to 2 mL)
Isolation from multiple sample types e.g. cell culture supernatant, urine, plasma, serum, ascites etc.

Western Blot analysis of NK cell EVs and platelet EVs

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Separation of EV subpopulations

Subpopulations of EVs can be separated using superparamagnetic MACS® MicroBeads. For example, NK cell EVs and platelet EVs can be separated using CD81⁺and CD9⁺MicroBeads.

MACS Handbook:

Tumor cells (human)

Tumor cells (mouse)

Magnetic cell separation

MACSPlex Analysis of plasma-derived extracellular vesicles (application note)

Related products:

Exosome Isolation Kits

Exosome analysis

For further downstream analysis of exosomes we have established the MACSPlex Exosome Assay. This multiplex bead-based assay consists of capture and detection antibodies to analyze the composition of exosome surface proteins in a given sample by flow cytometry.

Workflow of the multiplex bead platform

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Advantages of the MACSPlex Exosome Assay

Comprehensive – Fast and easy screening of 39 surface markers on EVs
Flexible – Additional antibodies can be added for multi-parameter analysis
Express Mode – Automated acquisition and analysis in combination with the Express Mode on the MACSQuant® Flow Cytometers.

Surface marker profile of EVs isolated from melanoma cell cultures or colon cancer cells

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Discrimination of EVs by differing tumor markers

EVs isolated from cancer cell lines can be discriminated by differing markers from tumor entities. EVs from melanoma cell line for instance show different surface markers then EVs from colon cancer cells as demonstrated by binding to the respective MACSPlex Exosome capture beads.