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    Plasma Exosome Isolation

    Silvia Cerri

    Published: 2022-09-16 DOI: 10.17504/protocols.io.81wgb7xnqvpk/v1
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    Abstract

    This protocol details methods for the isolation of small extracellular vesicles from plasma.

    Before start

    Keep samples On ice during the entire procedure!

    Steps

    Plasma Exosome Isolation

    1.

    Incubate plasma samples (starting volume: 1-1.5ml) On ice for complete thaw.

    2.

    Filter samples through a 13mm sterile syringe filters with a 0.8 µm Supor (PES) membrane.

    3.

    Centrifuge at 20000x g,4°C to remove large extracellular vesicles.

    4.

    Collect the supernatant and filter it through 13mm sterile syringe filters with 0.2 mm Supor (PES) membrane.

    5.

    Centrifuge at 100000x g,4°C to pellet small extracellular vesicles.

    6.

    Remove the supernatant, resuspend the pellet in 1mL and repeat the previous step (repeated again in the next step for convenience).

    Note
    This step is optional, according the final application of the pellet.

    7.

    Centrifuge at 100000x g,4°C to pellet small extracellular vesicles.

    8.

    Process the pellet according to the type of analysis.

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