Total Exosome Isolation Kit (from plasma) (Invitrogen™)

The Total Exosome Isolation Kit (from plasma) enables fast and efficient enrichment of intact exosomes from plasma samples. The kit consists of two parts: Total Exosome Isolation Reagent (from plasma) and Proteinase K (20 mg/mL). The Total Exosome Isolation Reagent enables simple and reliable concentration of intact exosomes from plasma samples with a protocol that is scalable depending on your sample size. Proteinase K is provided separately for optional upfront plasma treatment and is used to eliminate all plasma protein and thus maximize the purity of the exosome preparation.

• Maximize recovery of intact exosomes for any type of downstream application
• Easily isolate exosomes using our simple and reliable protocol
• Avoid time-consuming ultra-centrifugation
• Maximize flexibility—scale up or down depending on the sample size

By tying up water molecules, the Total Exosome Isolation Reagent forces less-soluble components such as vesicles out of solution, allowing them to be collected by a short, low-speed centrifugation. After a 10 minute treatment with Proteinase K, the reagent is added to the plasma and the solution is incubated for 30 min at 2–8°C. The precipitated exosomes are recovered by standard centrifugation at 10,000 x g for 5 min at room temperature. The pellet is then resuspended in PBS or similar buffer, and the exosomes are ready for downstream analysis or further purification by affinity methods. Total RNA and protein can then be purified using the Total Exosome RNA and Protein Isolation Kit (Cat# 4478545), or the intact exosomes can be used for biological studies of their pathways, functions, and trafficking.

For downstream mass spectrometry analysis we recommend performing gel electrophoresis purification of the isolated exosome suspension prior to analysis.