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2 best Reasons to use our kits to purify exosome


95%  exosome purity isolation

10 fold  higher exosome yield than all other kits

Our Exosome kits help avoid 80% Exosome waste in your sample.

 

Product Application & Flow Chart: (click on products to show/hide description)

 

P100 PureExo Cell: cell media exosome

P101 PureExo Serum: serum exosome

P120 DiagExo Urine: urine exosome

P121 DiagExo BodyFluid: body fluid exosome

P200 RNA/Protein Kit: exosome RNA / Protein

P300 Serum ExoProtein: serum Protein

P400 ExoFectin: small RNA into exosome

P401

P900

P701-5 Human Wnt-3a

P702-5 Mouse Wnt-3a

P703-5 Human Wnt-5a

P704-5 Mouse Wnt-5a

P801 Human Neural Stem Cell

P501 Total Protein Cell

P502 Total Protein Tissue

P503 Plasma Membrane Protein Extraction Kit

P505 Detergent-free Total Protein Extraction Kit


Our kits optimize your exosome research result.

Cat. #: P101 (10 reactions); P101L (40 reactions); P101S (2 reactions)

Storage: keep all bottles upright, in cool and dark place. Shelf Life: 12 months

Product Size: Each reaction can process 100~500 μL serum or plasma. The yield of each reaction is 100~200 μL exosome, from which 300~400 μg exosomal protein or 200~300 ng exosomal RNA can be extracted.

Product Description (This product is for research use only.)

This kit can isolate / purify pure exosome at high yield from serum or plasma.

  • Easy to use: No ultra-centrifugation (< 2 hours)
  • 10 fold higher yield (vs. other kits and ultracentrifuge)
  • Save cost (vs. antibodies-beads method)
  • Isolate Pure exosome (>95%)
  • Intact exosome (good morphology)

  • Use as little as 100 μL serum to achieve high yield of exosomes for any downstream applications: EM study, exosome label, exosome subpopulation, qRT-PCR profiling of exosomal miRNAs, and gel analysis of exosomal proteins.

    Product Contents (store in room temperature)

    ComponentAmount
    Cat. #: P101Cat. #: P101LCat. #: P101S
    Solution A2.5 mL10 mL0.5 mL
    Solution B2.5 mL10 mL0.5 mL
    Solution C2.5 mL10 mL0.5 mL
    Sample Buffer20 mL80 mL4 mL
    PureExo® Column10402

    * Tightly cap all bottles immediately after each use to prevent evaporation.

    Do not process more than 500 μL serum or plasma for each reaction. Otherwise it will cause indistinct layer separation and column clogging.

    Protocol (for processing 100-500 μL serum / plasma)

    1. Collect the serum or plasma sample and keep it on ice. If start with frozen sample, thaw the sample completely at room temperature, and keep it on ice.

    2. Centrifuge the serum / plasma sample at 3,000× g for 15 minutes at 4oC to remove debris.

  • Important: Skipping this step may cause filter clogging in step 15.
  • 3. Transfer 100 - 500 μL clear supernatant to a 15mL centrifuge tube without disturbing the pellet. Do not process more than 500 μL sample per reaction. Add Sample Buffer to make a total volume of 2 mL diluted serum / plasma sample, and keep it on ice. (This dilution works well for starting sample range of 100 - 500 μL.)

    4. In another 1.5 mL microcentrifuge tube, add Solution A/B/C in the following order to prepare 0.75 mL mixture A/B/C (always prepare mixture A/B/C right before use):

    1st add Solution A 0.25mL

    2nd add Solution B 0.25mL

    3rd add Solution C 0.25mL

    * Cap all bottles well immediately after each use to prevent evaporation.

    5. Vortex the mixture A/B/C for 10 seconds to obtain a homogenous solution.

    6. Add the 0.75 mL mixture A/B/C to the 2 mL diluted serum / plasma sample (from step 3).

    7. Cap the 15 mL tube, vigorously vortex for 30 seconds, then incubate at 4°C for 30 minutes.

    8. The mixture now appears as 3 layers:

    Pipet out the Top transparent layer and discard it without disturbing the Middle fluffy layer.

    9. Transfer the Middle fluffy layer (Exosome is in this layer) to another 1.5 mL microcentrifuge tube. Spin the tube at 5,000× g for 3 minutes. A new three-layer separation will appear: Top transparent layer, Middle fluffy layer and Bottom colorless layer. (See figure below.) Proceed to the next step immediately.

    Top transparent layer

    Middle fluffy layer (Exosome is in this layer.)

    Bottom colorless layer

    10. Pipet out the Top transparent layer and discard it. Insert pipette tip down to the tube bottom to completely remove the Bottom colorless layer. Only keep the Middle fluffy layer in the tube. Exosome is in this layer.

    11. Spin again at 5,000x g for 3 minutes, and 3 layers will appear again. Now, repeat step 10 for one more time. Now only the “fluff pellet” left in the tube. The “fluff pellet” volume is about 25 μL in our case.

    12. Leave the tube cap open to air dry for 5-10 minutes at room temp (do not over dry).

    13. Add 1× PBS equal to 4 times volumes of the collected fluff pellet to the tube. In our case, we added 100 μL PBS (4 x 25 μL fluff pellet). Resuspend the fluff pellet by pipetting up and down vigorously for 40 times.

    14. Shake the tube on a horizontal shaker at high speed for 3 minutes, then pipet up and down vigorously for 10 times. Repeat this “shake-pipet up and down” for another 2 times.

    Note: This step is important. If the fluff pellet is not well re-suspended, the exosome may be trapped in the fluff pellet resulting in low exosome purity and yield. Some types of samples, eg. Hyperlipidemia patient serum sample, it is difficult to dissociate the fluff pellet to release exosome. In such case, extend the pipetting and shaking time in step 13 and 14.

    15. Spin the tube at 5,000x g for 5 minutes. Without disturbing the “fluff pellet”, transfer the supernatant carefully into one PureExo® Column (provided).

    Note: Keep the “fluff pellet” at 4oC. Do not discard it until the experiment is finished. See “Trouble shooting” 2.2 for detail.

    16. Spin the PureExo® Column at 1,000× g for 5 minutes to collect all the “flow-through”.

    17. The “flow-through” is the isolated pure exosome (exosome suspended in PBS). The whole protocol is completed here. Use it directly for downstream assays (e.g. use 101Bio Exosomal RNA and Protein Extraction Kit, Cat.#: P200, to extract exosomal RNA/Protein), or store at 4°C for up to 1 week , or at ≤80°C for up to 3 months. Concentrated exosome will precipitate after sitting. Pipet up and down to resuspend it well before each use.


    More info: AAV - Adv Release Solution (10 mL, 20 Rxn)

     

    Supplier: 101Bio