Diagenode

ExoIP Composite Kit

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Catalog Number
Format
Price
C28010001
$490.00

ExoIP Composite Kit consists of immunoprecipitation method using a mixture of magnetic beads coupled with antibodies that recognize CD9, CD63, CD81 and EpCAM antigens for an optimal pure and high quality of exosome isolation. 

ExoIP Kit - Benefits

  • Highly pure exosome population: ideal for RNA and non-coding RNA analysis (microRNA) protein analysis, Western blot, ELISA, flow cytometry, and more!
  • Ready to use, high-quality: includes antibody-conjugated beads and optimized BSA-free buffers
  • Sample diversity: serum, plasma, and cell culture supernatant
  • Highly validated with multiple methods: Western blot, electron microscopy, RT-qPCR, dynamic light scattering, and flow cytometry

  • Exosomes

    Exosomes are small extracellular vesicules of 30-100 nm constitutively released under certain stimuli in biological fluids including blood, urine, saliva, breast milk, culture medium of cell cultures, etc.

    Exosomes contain various molecular constituents of their cell of origin, including proteins and microRNA. Circulant exosomes can potentially act as cargo messenger and transfer molecules from one cell to another via membrane vesicle trafficking.                                                                                                                                             .                                                                                                               Exosomes are suitable for a variety of downstream applications including microRNA analysis, protein analysis and functional use.

  • Product description

    Description

    Diagenode's ExoIP Composite kit was developed to optimize serum, plasma and cell culture supernatant exosome isolation.  

    This ready to use kit contains :
    • Mixture of magnetic capture beads coupled with anti-CD9, anti-CD63, anti-CD81 and anti-EpCAM antibodies that recognize exosome surface CD9, CD63, CD81 and EpCAM antigens.
    • Treatment buffer that decreases the non-specific binding on the magnetic bead
    • Washing/dilution buffer
    • Exosome elution buffer to recover after immunocapture fully intact and functional exosomes

    Validation data

    We used several methods to charachterize and validate exosome isolated with our immuno capture ExoIP Composite kit:

    • Validation by dynamic light scattering

    Characterization of vesicle population according their overall size by dynamic light scattering (from a Zetasizer Nano S, Malvern Instruments).
    Diagenode’s ExoIP yield to highly pure exosome preparations. The measured size of the vesicle population is in perfect accordance with the theoretical size range expected for exosomes: 30-100 nm.

     

    • Validation by electron microscopy

    The immunogold anti-CD9 labeling confirms the highly pure exosome preparations from HeLa Cell culture medium by showing clean exosomes in the samples.
    The vesicle highlighted herein is identified as an exosome according to their size and it characteristic morphology but also the precise anti-CD9 tag with the gold nanoparticles.

     

    • Validation by Western Blot on cell culture medium

    Western Blot results show that internal exosomal proteins Alix, HSP70 and TSG101 and membranous exosomal proteins CD63, CD9 and CD81 are expressed in ExoIP isolated vesicules from HEK 293T culture medium. These data demonstrate that vesicles isolated with the Diagenode’s ExoIP kits are very pure exosomes.

     

    • Validation by Western Blot on serum and plasma



    1. CD9 EoxIP kit
    2. CD63 ExoIP kit
    3. CD81 ExoIP kit
    4. Composite ExoIP Kit

     

    • Validation by Western Blot: comparison with competitor kits

    Western Blot results show that internal exosomal protein HSP70 is highly expressed in exosomes isolated from culture medium of HEK 293T with ExoIP Kits compared to competitor immunocapture bead kits. WCL= whole cell lysate.

    • Validation by flow cytometry

    Flow analysis performed on isolated exosomes from HEK 293T cell culture supernatant with ExoIP CD9 kit (BD FACS Canto II). The amount of input (ml) and beads are equal in each tested condition.

    Flow analysis performed on isolated exosomes from different volumes of HEK 293T cell culture supernatant with ExoIP CD81 kit (BD FACS Canto II). Significant signal is obtained from low sample input. Signal is increasing according to the starting volume, reflecting the important binding capacity.

    Example of downstream application data

    qPCR application analysis to detect miRNA

    Relative miR-21 mRNA expression detected by qPCR on plasma exosomes isolated with ExoIP kits. No detection by ultracentrifugation exosome isolation.

  • Documents
    ExoIP kits MANUAL
    ExoIP kits are the appropriate solution to isolate and enrich exosomes from diverse biological fl...
    Download
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: ExoIP Composite Kit (Diagenode Cat# C28010001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

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