Kit Components
| Item |
Quantity |
Storage Temperature |
| Total Exosome Isolation Kit (Cell Culture Media) |
60 ml/120 ml* |
+4oC |
| ExoSafe™ Buffer |
2 ml |
-20oC |
* Sufficient for processing 120/240 ml of cell culture media.
Product Overview
Extracellular vehicles (EVs) such as exosomes are nano size vesicles ranging from 30-150 nm which are secreted by all living cells. They are present in almost all biological fluids. EVs (exosomes) carry lipids, proteins, and RNAs derived from their parent cells, and serves as messengers of intracellular network. EVs can also be a valuable source for biomarker discovery.
Currently, the isolation of high-quality EVs (exosomes) from various types of samples is still challenging. While differential ultracentrifugation is widely used for the isolation of exosomes, it requires the use of special equipment with several drawbacks such as its time-consuming operation and unable to process a large volume of samples.
Exom Biopharma provides an easy solution for exosome isolation that can help you to enrich and purify high-quality, intact exosomes from a variety of biological fluids, including cell culture media, serum, plasma, urine, saliva, milk, and plants. The Exosome Isolation Kit (cell culture media) is not only simple, fast, and easy for use, but also can clean up exogenous nucleic acid contaminations and stabilize exosomes for downstream biomarker analysis and drug delivery application. Our Total Exosome Isolation kit from cell culture media is based on simple precipitation regent. Along with the kit we also provide exosome resuspension buffer which will prevent the exosomal RNA and proteins from degradation. The process of exosome isolation is simple and easy:
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Advantages of our Kits
- High purity and yields
- Includes ExoSafe™ Exosome Storage Buffer that contains stabilizing chemicals for long term storage of the exosomal proteins and RNA.
Applications
Isolated extracellular vesicles can be used for the following application:
- Packaging drugs, small molecules, and siRNA
- Added as is to the cells for expression analysis
- WB, and PCR and RT-PCR applications
General Guidelines
- Do not grow cells in normal fetal bovine serum because the serum contains its own exosomes. The cells should be grown in either serum free media or media with our Exosome Depleted Fetal Bovine Serum (Catalog # E2003).
- Our ExoSafe™ Exosome Storage Buffer (Catalog # E2002) contains stabilizing chemicals for long term storage of the exosomes.
- Store isolated exosomes at -20oC or -80oC for long term storage.
Protocol
- Harvest cells by centrifugation at 3,000xg for 10 minutes and transfer supernatant containing exosomes in a fresh tube.
- Centrifuge Cell Culture Media at 10,000x g for 20 minutes.
- Transfer supernatant to a fresh tube. Discard the pellet.
- Add ExoPrep™ Reagent (from cell culture media) reagent as follows:
| Culture Media |
ExoPrep™ Reagent |
|
1 ml
|
0.5 ml
|
|
5 ml
|
2.5 ml
|
|
50 ml
|
25 ml
|
- Invert tube or vortex to mix the contents well.
- Incubate the contents at 2-8oC for 2 hours or for maximum yield incubate the mixture overnight.
- To collect the exosome, centrifuge the tubes at 10,000x g for 30 minutes at 2-8o
- Aspirate or discard supernatant. Exosomes will be visible as solid pellet in the bottom of the tube.
- Carefully remove and discard the supernatant and save the pellet. Re-centrifuge the tube for 1 minute at max speed.
- Carefully remove any excess supernatant.
- Resuspend exosome pellet in ExoSafe™ buffer (200 µl for 1ml Cell Culture or 500 µl for 5 ml cell culture supernatant)
- Optional: To remove exogenous RNA/DNA, add 1µl or 2.5 µl of Exonuclease Plus to the resuspended exosomes and incubate at 37oC for 20 minutes followed by 50oC for 10 minutes to inactivate the ExonucleasePlus™.
- Centrifuge the tube again at 10,000xg for 10 minutes. Transfer the supernatant that contains the exosomes into a fresh tube and store isolated exosomes at -20oC (for short term) or -80oC for long term stability of the exosomes.
