Exosome Isolation and Purification Plus Kit (From Plasma and Serum) 

Exosomes are small vesicles (30 - 150 nm) containing RNA and protein that are secreted by various types of cells in culture, and found in abundance in body fluids including blood, saliva, urine, and breast milk. Exosomes are thought to function as intercellular messengers, delivering their cargo of effector or signaling macromolecules between specific cells.

MCE Exosome Isolation and Purification Plus Kit (From Plasma and Serum) provides a simple and effective method to isolate and purify intact exosomes from plasma and serum that can be used for electron microscope analysis, NTA analysis, WB, qPCR , cell biology studies, and animal experiments.

Solution A1: -20°C, 2 years.

Solution B1, Exosome Purification Filter (EPF): RT, 2 years.

1. During sample preparation, if the sample concentration is too high or too low, appropriate dilution or concentration can be performed. Centrifugation can be repeated until no significant pellet is observed.

2. The products in this kit do not contain RNase or DNase. During the experiment, avoid introducing RNase and DNase contamination.

3. To ensure that the exosomes obtained are from your cells, use exosome-free serum to culture the cells.

4. It is recommended to use freshly prepared samples and avoid repeated freeze-thaw cycles.

5. If you need to co-culture the obtained exosomes with cells later, you can filter them through a 0.22 μm filter membrane to remove bacteria.

6. This product is for R&D use only, not for drug, household, or other uses.

7. For your safety and health, please wear a lab coat and disposable gloves to operate.

1. Experiment Preparation

1) Pre-cool the centrifuge at 4°C for 10 min before use.

2) Place Solution A1 in a 4°C refrigerator to thaw, then take an appropriate amount of Solution A1 and keep it on ice for later use.

Note: It is recommended to aliquot the Solution A1 after the first thaw to avoid repeated freeze-thaw cycles.

2. Sampling: For frozen samples, thaw them in a 25°C water bath and then place on ice for use. For fresh samples, directly place on ice for use.

3. Sample Aliquoting: Aliquot the sample into 500 μL/tube. If the sample is less than 500 μL, supplement with PBS (1×).

4. Centrifugation to Remove Impurities

1) Centrifuge at 4°C, 3,000 g (6,200 rpm) for 10 min, and collect the supernatant.

Note: If a large amount of precipitate is observed, repeat the centrifugation until no significant precipitate is present.

2) Centrifuge at 4°C, 12,000 g (12,400 rpm) for 10 min, and collect the supernatant.

Note: The above centrifugation steps are based on a small centrifuge (effective radius ~ 7 cm) and ≤ 2 mL centrifuge tubes. For larger systems, it is recommended to centrifuge in separate tubes, the same applies below.

1. Add 400 μL of pre-cooled Solution A1 to each 500 μL of the previously prepared sample, immediately seal the centrifuge tube, and vortex for 30 s to mix thoroughly.

2. Centrifuge at 4°C, 12,000 g (12,400 rpm) for 20 min, and collect the supernatant.

1. Add 120 μL of Solution B1 to every 500 μL of the sample, immediately seal the centrifuge tube, and vortex for 1 min to mix thoroughly. Incubate at 4°C for 30 min.

Note: Extending the incubation time may increase exosome yield, but do not exceed 24 h.

2. Centrifuge at 4°C, 12,000 g (12,400 rpm) for 15 min, and collect the pellet, which is rich in exosome particles. Carefully remove the supernatant completely.

Note: The pellet may be hard to observe. It is recommended to mark the pellet position before centrifugation.

3. Centrifuge at 4°C, 12,000 g (12,400 rpm) for 2 min, and collect the pellet.

4. Resuspend the exosome pellet in an appropriate amount of PBS (1×) to dissolve it thoroughly.

Note: It is recommended to resuspend the exosome pellet in 200 μL PBS for every 500 μL of sample.

5. Centrifuge at 4°C, 12,000 g (12,400 rpm) for 2 min, and collect the supernatant. The supernatant contains the exosome particles.

Note: If a large amount of pellet is observed, repeat the centrifugation until there is no visible pellet.

1. Exosome Purification

Transfer the collected crude exosomes to the exosome purification filter’s upper chamber, and centrifuge at 4°C, 3,000 g (6,200 rpm) for 10 min. Collect the liquid at the bottom of the filter tube, which contains the purified exosomes.

2. Exosome Storage

Aliquot the purified exosomes and store at -80°C for future use. Avoid repeated freeze-thaw cycles.

1. During sample preparation, if the sample concentration is too high or too low, appropriate dilution or concentration can be performed. Centrifugation can be repeated until no significant pellet is observed.

2. The products in this kit do not contain RNase or DNase. During the experiment, avoid introducing RNase and DNase contamination.

3. To ensure that the exosomes obtained are from your cells, use exosome-free serum to culture the cells.

4. It is recommended to use freshly prepared samples and avoid repeated freeze-thaw cycles.

5. If you need to co-culture the obtained exosomes with cells later, you can filter them through a 0.22 μm filter membrane to remove bacteria.

6. This product is for R&D use only, not for drug, household, or other uses.

7. For your safety and health, please wear a lab coat and disposable gloves to operate.