Exosome-Human EpCAM Isolation Reagent (from cell culture) - FAQs

Yes. See this poster (https://tools.thermofisher.com/content/sfs/posters/Exosome-poster-ISEV-2013-Boston.pdf).

In addition, here are some citations:
- Blood 91:2573 (1998)
- Science 289:444 (2000)
- J Physiol 537:537 (2001)
- Mol Cell Proteomics 12:587 (2013)
- Biol Reprod 81:717 (2009)

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Yes, exosomes isolated with different surface markers can be distinctive in their protein profile. This has been demonstrated by Tauro et al. (http://www.ncbi.nlm.nih.gov/pubmed/23230278), who isolated two distinctive populations of exosomes based on surface markers EpCam or A33 from conditioned cell culture medium from a human carcinoma cell line. This proteomics study indicated that these two populations of exosomes are unique.

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We have exosome isolation kits for Exosome-Human CD63 (Cat. No. 10606D), Exosome-Human CD9 (Cat. No. 10614D), Exosome-Human CD81 (Cat. No. 10616D), and Exosome-Human EpCAM intended for isolating exosomes with these commonly used exosome surface antigens. If you are interested to isolating exosomes with other specific surface markers using your own antibody, you can use our Dynabeads exosome immunoprecipitation (Protein A, Cat. No. 10610D), Dynabeads exosome immunoprecipitation (Protein G, Cat. No. 10612D), or Exosome-Streptavidin for isolation/detection (Cat. No. 10608D). In addition, anti-mouse IgG Dynabeads magnetic beads (Cat Nos. 11031 or 11033) also can be employed in exosome isolation using mouse monoclonal antibodies against selected surface markers.

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Exosomes are usually characterized by flow cytometry (using surface markers such as CD9, CD63, TSG101, and Alix), by EM to study morphology and size, or by detailed protein analysis by LC-MS/MS.

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It depends on the cell source from which the exosomes were derived. The most commonly used surface markers for isolating and characterizing exosomes are CD9, CD63, CD81, or TSG101. Here are some of the recent references and surface markers for identifying or isolating exosomes:

Alix, CD63, EpiCam, HSP70, TSG101 Mol Cell Proteomics 12:587 (2013)
CD9, CD63 Hum Mol Genet 21:R125 (2012)
CD63, MHC IIJ Biol Chem 278:52347 (2003)
CD9, CD81, Lamp1, TSG101 Cancer Res 67:7458 (2007)
CD63 Nature Cell Biol 9:654 (2007)
Alix, CD37, CD53, CD63, CD81, CD82, TSG101J Cell Biol 200:373 (2013)
CD59, CD63, CD133, TSG101 FASEB J 23:1858 (2009)

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Exosomes can be isolated by ultracentrifugation or density gradient separation in addition to a precipitation approach. Exosomes can also be isolated by a magnetic approach using Dynabeads magnetic beads targeting exosome markers such as Human CD9, CD63, CD81, EpCAM or secondary antibody-coated Dynabeads magnetic beads using different antibodies against other exosomal surface markers.

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A range of different functions have been reported such as antigen presentation, apoptosis, angiogenesis, inflammation, and coagulation by protein/lipid exchange or activation of a signaling pathway. Exosomes provide a novel vehicle for genetic exchange between cells and mediate cell to cell communication. Exosomes also transport and propagate of infectious cargoes such as prion and retrovirus.

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Exosomes are small, membrane-bound ovoid to cup shaped particles around 30-150 nm in size containing mRNA, microRNA, proteins, and lipids. Exosomes are released by normal, abnormal, or neoplastic cells into body fluid such as blood, urine, saliva, and breast milk. Exosomes originate from the endocytic compartment and are released from cells as multivesicular bodies (MVB) fused with plasma membrane (J Cell Biol 200:373 (2013)).

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