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Invitrogen™ Exosome-Human EpCAM Isolation Reagent (from cell culture)

Catalog No. 10618D
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10618D 2 mL
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Description

Obtain a highly pure exosome subset with easy protocol scalability with Exosome-Human EpCAM Isolation Reagent (from cell culture).

Obtain a highly pure exosome subset with easy protocol scalability with Exosome-Human EpCAM Isolation Reagent (from cell culture). Following isolation, the specific sub-population of exosomes, ECVs can be further studied using methods such as Western blotting, qPCR, or next generation sequencing.

  • Minimal hands-on time for specific exosome isolation
  • Compatible with most downstream analysis
  • Recent studies suggest that exosome/ECV population consists of several subsets, possibly with different functions
  • Using immuno-affinity based Dynabeads magnetic separation technology, these subsets can be selectively isolated and studied
  • Cell culture media must first be enriched for exosomes
  • Can be done quickly and efficiently using Total Exosome Isolation Reagent (from cell culture media) or traditional ultracentrifugation
  • Following enrichment, specific exosomes are isolated by binding to surface of magnetic beads
  • In addition to their high-quality sensitivity, reproducibility, and stability, Dynabeads allows you to see samples during handling due to color of magnetic beads
  • When a sample tube is placed in the magnet, bead-bound exosomes are pulled to side of tube, allowing for easy separation and buffer changes
  • Sample and bead volume can be easily scaled according to your sample size or downstream applications

Mix thoroughly to ensure that magnetic beads do not settle in the sample tube. A mixer which provides both tilting and rotation for optimal mixing is recommended.

Specifications

Content And Storage Store in refrigerator (2–8°C).
Product Type Isolation Reagent
Product Line Dynabeads
Quantity 2 mL
Sample Type Cell Cultures

Frequently Asked Questions (FAQs)

Do you have any references or citations using Dynabeads magnetic beads to isolate exosomes?

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)

Can exosomes isolated with different surface markers be different?

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.
What kind of product do you suggest if I would like to use Dynabeads magnetic beads to isolate exosomes?

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.

How are exosomes characterized?

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.
What are the commonly used surface markers for isolating exosomes?

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)

How are exosomes isolated?

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.
What are the functions of exosomes?

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.
What are exosomes?

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)).

For Research Use Only. Not for use in diagnostic procedures.