Exosomes are small extracellular vesicles, typically 30–150 nm in diameter, released by a wide range of cell types. They carry diverse molecular cargo—including RNA species, proteins, and phospholipids—and are abundant in cell culture supernatants as well as biological fluids such as milk, serum, plasma, semen, saliva, urine, amniotic fluid, and cerebrospinal fluid. By mediating the transfer of molecular information between cells, exosomes play key roles in intercellular communication and regulation.
This kit provides a straightforward and reliable method for isolating intact exosomes from cell culture supernatants and various biofluids, including serum, plasma, breast milk, urine, and saliva. The resulting vesicles are suitable for a broad range of downstream applications, such as electron microscopy, nanoparticle tracking analysis (NTA), NanoFCM particle characterization, Western blotting, quantitative PCR, and high throughput sequencing.
This product should be stored at 2-10°C up to 12 months. The expiration date is indicated on the outer label of the kit box.
Sample Type
Protocol
Cell Culture Supernatant
Collect culture supernatant, centrifuge at 1500 g for 10 min, then concentrate to ~0.5 mL using an ultrafiltration tube. Filter the concentrated sample through a 0.45 µm filter before isolation.
Serum, Plasma, Breast Milk, Saliva
Centrifuge 1 mL of sample at 10,000 g for 10 min and collect the supernatant. Filter through a 0.45 µm filter and use up to 0.75 mL for isolation.
Urine
Collect 10–50 mL first-morning midstream urine, centrifuge at 1500 g for 10 min, and concentrate the supernatant to 0.5 mL. Filter through a 0.45 µm filter before isolation.
Plant
Extract 35 mL plant juice, incubate at 4 °C for 1 h, then centrifuge at 10,000 g for 10 min. Concentrate the supernatant to 0.5 mL and filter through a 0.45 µm filter before extraction.
Step
Protocol
1
Mix 150 µL Reagent A with 1 mL Reagent E in a 1.5 mL tube, invert to mix, centrifuge (850 rpm, 1 min), discard supernatant and keep the precipitate.
2
Dissolve Reagent B in 2000 µL sterile water, transfer 1000 µL to the precipitate in step 1 and incubate on a level mixer at room temperature for 2 h.
3
Centrifuge at 850rpm for 1min, aspirate the liquid and retain the precipitate.
4
Add 1000ul of Reagent E, centrifuge at 850rpm for 1min, aspirate the liquid and retain the precipitate. Repeat this step twice.
5
Add the sample, then Reagent C and Reagent D in a 125:1:125 ratio (Reagent:C:D) and incubate overnight at 4 °C (or 2 h at room temperature with mixing).
6
Centrifuge at 850rpm for 1min, aspirate the liquid and retain the precipitate.
