Exosomes are small vesicles (30–150 nm) containing sophisticated RNA and protein cargos. They are secreted by all cell types in culture and are found to occur naturally in body fluids, including blood, saliva, urine, CSF, and breast milk. The precise molecular mechanics for their secretion and uptake, as well as their composition, “cargo”, and resulting functions, are only beginning to be unraveled. Exosomes are now viewed as specifically secreted vesicles that enable intercellular communication and have become the focus of exponentially growing interest, both to study their functions and to understand ways to use them in the development of minimally invasive diagnostics.

We offer an array of reagents, tools, and protocols for isolation, characterization, and analysis of their RNA and protein content, as well as in vitro and in vivo tracing to help you further the understanding of exosomes.

Exosomes FAQs

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Products for exosome research

Total exosome isolation

Ultracentrifugation can be tedious and time consuming. Now you can easily enrich for intact exosomes from a variety of starting samples using the flexible and scalable Total Exosome Isolation reagents. These reagents and accompanying protocols are ideal for a range of experimental set-ups including low volume input and handling of multiple samples.

Isolation of exosome subpopulations

Total exosomes enriched from cell culture (by using the Total Exosome Isolation reagents or ultracentrifugation) can be further purified into specific subpopulations by immunomagnetic capture. Use the Dynabeads®-based CD9, CD63, CD81, or EpCam-specific reagent, or combine the Streptavidin reagent with your choice of a biotinylated antibody to purify any population based on a surface antigen.

Visual identification of exosomal vesicles can be challenging, and free exosomes alone are too small to be detected in flow. There are a few methods, though, which can be used for analysis.

  • NanoSight® Instrument - Exosomes recovered with the Total Exosome Isolation reagent can be analyzed using the NanoSight® instrument for approximate size range and concentration.

  • Light Microscopy - Exosomal RNA or membrane components can be labeled, allowing for visualization of exosomes under the microscope. Specialized spin columns are available to remove unincorporated dyes. These columns also allow for buffer exchange and easier desalting or removal of low molecular weight (< MW 3000) contaminants compared to current methods.

  • Electron Microscopy (EM) - One of the advantages of using Dynabeads® magnetic separation technology to pull out pure and specific exosomes is that you can move directly to EM analysis. You can analyze subsets from total exosomes enriched from cell culture using CD9, CD63, CD81, or EpCam-specific reagent, or use the Streptavidin reagent in combination with your choice of biotinylated antibody.

  • Flow Cytometry - Immunomagnetic capture of exosomes on the surface of the Dynabeads® allow for a clear and defined FFC/SSC detection typically in less than one hour. You can analyze subsets from total exosomes enriched from cell culture using the CD63-specific reagent, or use the Streptavidin reagent in combination with your choice of biotinylated antibody.

Exosomes have been shown to transport a range of molecules from one cell to another. Their cargo includes proteins, lipids, mRNA (fragments and full length), rRNA, miRNA, and various ncRNA.

  • RNA & Protein Isolation - Simultaneous isolation of both proteins and total RNA from the same sample of pre-enriched exosomes can be achieved using the Total Exosome RNA and Protein Isolation Kit. The kit is compatible with all protocols for exosome isolation.

  • Protein Analysis - Specific monoclonal antibodies (CD9, CD63 and CD81) allow for detection of cellular and exosomal antigens by Western blot analysis. If you are looking to compare multiple protein samples on the same gel and want to keep exosomal protein complexes intact, you should check out the fast and gentle Dynabeads®-based immunoprecipitation method using Protein A or Protein G in combination with your primary antibody of choice.

  • RNA Analysis - Exosomal RNA recovered using the Total Exosome RNA and Protein Isolation Kit can be analyzed e.g. by qRT-PCR using TaqMan® assays, RNA sequencing, or next generation sequencing tools. Exosomal RNA and membrane components can be labeled using fluorescent dyes, while unincorporated dye can be removed by using dedicated spin columns.

Gibco® Exosome-Depleted FBS significantly outperforms existing exosome-depleted FBS products on the market.

  • ≥90% of exosomes depleted
  • Cell culture testing of every lot
  • Specially developed for exosome research

Learn more about Gibco® Exosome-Depleted FBS

Documentary mini-series: "Exosomes-The Next Small Thing"

References

These published articles all cite the use of our exosome-specific products:

  • Expression of B-Cell Surface Antigens in Subpopulations of Exosomes Released From B-Cell Lymphoma Cells
    Oksvold et. al. View abstract
  • MicroRNA-193b is a regulator of amyloid precursor protein in the blood and cerebrospinal fluid derived exosomal microRNA-193b is a biomarker of Alzheimer's disease
    Liu CG et.al. View abstract
  • Hepatocellular Expression of B-Cell Surface Antigens in Subpopulations of Exosomes Released From B-Cell Lymphoma Cells
    Oksvold MP et. al. View Abstract
  • Expression of Serum Exosomal MicroRNA-21 in Human Hepatocellular Carcinoma
    Wang H et. al. View Abstract
  • Programmed Cell Death 4 and MicroRNA 21 Inverse Expression Is Maintained in Cells and Exosomes From Ovarian Serous Carcinoma Effusions
    Cappellesso R et. al.  View Abstract
  • Exosomes from Drug-Resistant Breast Cancer Cells Transmit Chemoresistance by a Horizontal Transfer of MicroRNAs
    Chen W-x et. al. View Abstract
  • miR-145 suppresses thyroid cancer growth and metastasis and targets AKT3
    Boufraqech M et. al. View Abstract
  • Natural Killer Cell-Mediated Shedding of ULBP2
    Wang R et. al. View Abstract
  • Histone Deacetylase 3 Unconventional Splicing Mediates Endothelial-to-mesenchymal Transition through Transforming Growth Factor β2
    Zeng L et.al. View Abstract
  • Methods for the extraction and RNA profiling of exosomes
    Zeringer E et. al. View Abstract
  • Delivery of Functional Anti-miR-9 by Mesenchymal Stem Cell–derived Exosomes to Glioblastoma Multiforme Cells Conferred Chemosensitivity
    Munoz JL et.al. View Abstract
  • The Complete Exosome Workflow Solution: From Isolation to Characterization of RNA Cargo
    Schageman J et. al. View Abstract
  • Characterization of a Stem-like Subpopulation in Basal-like Ductal Carcinoma in Situ (DCIS) Lesions
    Li Q et.al. View Abstract
  • Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials
    Vlassov AV et. al. View Abstract
  • Tumor exosomes induce tunneling nanotubes in lipid raft-enriched regions of human mesothelioma cells
    Thayanithya V et.al. View Abstract
  • Systemically Circulating Viral and Tumor-Derived MicroRNAs in KSHV-Associated Malignancies
    Chugh PE et.al. View Abstract
  • Differences in exosome populations in human breast milk in relation to allergic sensitization and lifestyle
    Torregrosa Paredes P et.al. View Abstract
  • Analysis of the RNA content of the exosomes derived from blood serum and urine and its potential as biomarkers
    Li M et. al. View Abstract
  • Direct Isolation of Exosomes from Cell Culture: Simplifying Methods for Exosome Enrichment and Analysis
    Li M et. al. View Abstract
  • Muscle Releases Alpha-Sarcoglycan Positive Extracellular Vesicles Carrying miRNAs in the Bloodstream
    Li M et. al. View Abstract