Exosome Research Products

Exosomes are small vesicles (30–150 nm) containing sophisticated RNA and protein cargos. 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 Invitrogen Dynabeads products and other 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.

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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 Invitrogen 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.

exosome-isolation-fig1

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.

exosome-isolation-fig2

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 a 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. Invitrogen 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 Invitrogen 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 Invitrogen CD9, CD63, CD81, or EpCam-specific reagent, or use the Invitrogen 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 Invitrogen 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 Invitrogen 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 Applied Biosystems 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 Invitrogen spin columns.
Gibco Exosome-Depleted FBS significantly outperforms existing exosome-depleted FBS products on the market.

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

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 Invitrogen 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.

exosome-isolation-fig1

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.

exosome-isolation-fig2

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 a 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. Invitrogen 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 Invitrogen 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 Invitrogen CD9, CD63, CD81, or EpCam-specific reagent, or use the Invitrogen 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 Invitrogen 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 Invitrogen 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 Applied Biosystems 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 Invitrogen spin columns.
Gibco Exosome-Depleted FBS significantly outperforms existing exosome-depleted FBS products on the market.

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"

Resources

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