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Exosomes are membrane-secreted vesicles that carry complex nucleic acids, proteins, lipids, and metabolites. Exosomes play key roles in cell-to-cell communication and signal transduction. Because they transport and exchange a variety of cargo between cells, exosomes are used as non-invasive biomarkers for several diseases.
Exosomes are small (30–150 nm) vesicles containing DNA, RNA, proteins, lipids, and metabolites. Exosomes are constantly secreted by cells in vitro and in vivo. The precise molecular mechanics for their secretion and uptake as well as the composition of their cargo, and resulting functions, are still being discovered. Exosomes have become the focus of growing interest both to study their functions and to understand ways to potentially use them as treatments for diseases.
Exosomes’ primary function is to mediate near and long-distance intracellular communication, by transporting their cargo of nucleic acids, proteins, lipids, amino acids, and metabolites, reflecting their cell of origin. Exosomes are being investigated for the key roles they play in the following processes:
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Exosome isolation using ultracentrifugation is considered a traditional method of isolation that can be both time-consuming and can damage the exosomes, changing their morphology and functional properties. Exosomes can be separated from cells and heavy artifacts by tying up water molecules via polymer precipitation, thereby reducing the solubility of exosomes and bringing about their precipitation. Exosomes are then harvested following low-speed centrifugation. Total Exosome Isolation reagents and kits utilize this method and help speed up the process of exosome isolation beginning with sample collection—from cell culture media, serum, plasma, urine, or other body fluids—and ending with purified exosomes ready for downstream molecular analyses (Figure 1). The Total Exosome Isolation kits and reagents, and accompanying protocols, are ideal for a wide range of experiments, including processing low sample volumes and handling multiple samples.
Figure 1. Traditional method of exosome isolation via ultracentrifugation.
Exosome isolation using immunomagnetic separation is a method particularly suited to purify exosome subpopulations. Exosome-specific antibodies are bound to superparamagnetic beads for more efficient purification of exosomes. Exosome-Streptavidin Isolation/Detection Reagent helps isolate and detect exosomes when combined with a biotinylated primary antibody. These exosomes can then be detected using assays such as flow cytometry, electron microscopy, and western blot (Figure 2).
Figure 2. Another method of exosome isolation via immunomagnetic separation.
Antibody-specific exosome isolation reagents are also available to isolate exosome subpopulations. These antibodies include:
Following isolation, the specific subpopulation of exosomes can be further studied using methods such as western blot, qPCR, or next-generation sequencing.
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Detection and analysis of exosomes can be very challenging due to their small size (30–150 nm). However, there are a few methods used for exosome detection and analysis.
Exosome subpopulations can be defined by their surface receptor. Antibodies specific to these surface receptors are used to detect exosome subpopulations. The following antibody-specific exosome analysis reagents for exosome subpopulations include:
Following isolation, the specific subpopulation of exosomes can be further studied using methods such as western blot, qPCR, or next-generation sequencing.
View all exosome detection and analysis products
Exosomes have been shown to transport a range of molecules between different cell types. Their cargo includes various proteins, mRNA (fragments and full length), rRNA, tRNA, miRNA, and ncRNA.
Antibody | Type of exosome analysis |
---|---|
CD9 | Flow cytometry, western blot, qPCR, sequencing |
CD63 | Western blot, qPCR, sequencing |
CD81 | Flow cytometry, western blot, qPCR, sequencing |
EPCAM | Flow cytometry, western blot, qPCR, sequencing |
By using exosome-depleted FBS, the inefficiencies that come with ultracentrifugation of serum are lessened. Gibco exosome-depleted FBS has the highest level of exosome depletion compared to homebrewed material and competitor FBS while retaining the vital nutrients necessary for cell culture. Features of this specialty FBS include:
KingFisher instruments automate the isolation of DNA, RNA, proteins, exosomes, viruses, and cells. KingFisher instruments help eliminate cross contamination, produce reproducible results, and cut your hands-on time down to 40 minutes. Set up plates, press start, and walk away.
Learn more about KingFisher instruments for automated protocols
In this documentary mini-series, we asked ten prominent scientists to share their thoughts on exosome research.
For Research Use Only. Not for use in diagnostic procedures.