Exosome Extraction and Purification Kit (Serum-Plasma)

Exosomes are small vesicles (30-150 nm) containing RNA and protein secreted by cells, which are abundant in serum/plasma. Exosomes are considered to have the function of the inter-cellular messenger, transmitting their effectors or signal molecules between specific cells.

In the biological functional study of exosome, it is necessary to separate its complete particles. The Rapid Extraction Kit of Serum/Plasma Exosome independently developed by Umibiois suitable for the extraction of exosome in the serum/plasma after the optimization of the components. It helps to obtain high-purity exosomal particles rapidly and efficiently.

High-speed centrifuge (with rotation speed up to 12000 rpm), vortex oscillator; 1.5 mL centrifugal tube, PBS buffer solution, ice machine.

* Nuclease-free, Sterile

• · Covers cell supernatant, tissue, urine, serum, plasma, milk, etc.
• · High sample processing capacity, up to 5 times that of similar products.
• · Exosomes can be used for cell co-culture and omics research.
• · Dedicated to continuous optimization of exosome kits for research and development.
• · Trusted by over 300 customers, ensuring worry-free purchasing and use.
• · Cost-effective to support large-scale research while saving costs.

I. Sample Pretreatment

1. The centrifuge shall be pre-cooled for 10 min at 4℃ before use;

2. Sampling: in case of a frozen sample, thaw it at 2-8℃ after taken out from the fridge; in case of a fresh sample, store it at 2-8℃ after collected.

3. Sub-package the sample according to 500 μL /tube, and make up the insufficient part to 500 μ L with 1 × PBS.

II Impure Protein Removal

1. After taking out Solution A from the kit, place it in the freezer layer of the fridge for at least 1 hour. Take out the pre-cooled Solution A and place it on brash-ices;

2. Adding Solution A: add 400μL pre-cooled Solution A into 500μL blood sample and tightly cover the centrifugal tube immediately, and fully mix it using the vortex oscillator for 30 seconds;

3. Centrifuging to remove the protein: centrifuge the evenly mixed sample at 4℃ at 12000 rpm for 20 min to remove the impure protein in the sample;

4. Transferring the supernatant: transfer the centrifuged supernatant whose impure proteins have been removed into a new 1.5mL centrifugal tube;

III Exosome Extraction

1. Adding Solution B: add 120μL Solution B into the centrifuged supernatant whose impure proteins have been removed;

2. Mixing the solution: after adding Solution B, cover tightly the centrifugal tube and mix it fully using the vortex oscillator for 2 min. Then let it stand for 5 min at room temperature;

3. Precipitating the exosome: centrifuge the centrifugal tube with the mixed solution at 4℃ at 12000 rpm for 15 min and discard the supernatant;

4. Centrifuging again: centrifuge the centrifugal tube with precipitation at 4℃ at 12000 rpm for 2 min and discard the supernatant (note: the supernatant shall be sucked as much as possible). Leave the 1.5 mL centrifugal tube open at room temperature for 10 min;

5. Resuspending the exosome: take 200μL of 1×PBS and spin the centrifuged precipitate evenly. After it is dissolved, transfer the resuspended solution into a new 1.5 mL centrifugal tube;

6. Obtaining exosomal particles: centrifuge 1.5 mL centrifugal tube with resuspended solution at 4℃ at 12000 rpm for 2 min and keep the supernatant. This supernatant is rich in exosomal particles.

(Note: at this point, there may be a large amount of precipitates, which is a normal phenomenon)

IV Exosome Purification

1. Purifying the exosome: transfer the obtained crude product of exosomal particles into the upper chamber of Exosome Purification Filter (EPF column) and centrifuge at 4℃ at 5600 rpm for 20 min. After the centrifugation, collect the fluid at the bottom of the EPF column. This fluid is the purified exosomal particles; (Note: EPF columns are not reusable)

2. Storing the exosome: purified exosomes shall be sub-packed at 50-100μL and stored in a low-temperature fridge at -80℃ for subsequent experimental use.

V Storage Conditions

After receiving the product, separately store the reagents. Please mix it thoroughly before use.

Solution A: transport at room temperature and store at -18℃ after the kit is opened (the freezer layer of a conventional fridge);

Solution B: store at room temperature;

EPF column: store at room temperature;

V Precautions

This product is for life science researches only!

VI Other Items

For more technical questions, please contact Umibio for further answers.