Extracellular vesicles (EVs), a family of membrane enclosed vesicles secreted by various types of cells, are involved in cell-to-cell communication by transferring cargos, such as proteins, metabolites, and nucleic acids. A key caveat to the clinical exploitation of EVs is that traditional method of choice for EV isolation, the ultracentrifugation, requires prohibitively long time and efforts, this is even true for large scale studies and less experienced researchers who want to isolate EVs in a timely and cost-effective manner with a required yield and at a suitable scale. Therefore, a methodology that will allow the rapid isolation of EVs with high yield would greatly facilitate such applications.
One familiar method that is increasingly being applied to isolate EVs is the use of precipitants such as polyethylene glycol (PEG) combined with centrifugation to obtain pellet EVs for subsequent processing. Precipitation method has several advantages; (i) familiar to use, (ii) uses common laboratory equipment, (iii) suitable for both small and large sample volumes, (iv)high yield. However, it also has the limitations: (i) polymeric contaminants, (ii) extended processing time, (iii) complicated clean-up steps, (iv) affecting downstream analysis and quantification.
ExoPRISMTM EV Isolation Kit, as a novel precipitation method, is introduced to overcome those hurdles by using low-molecular-weight electrolytes instead of the PEG polymer. (i) The isolated and enriched EV sample by ExoPRISMTM doesn't have polymeric contaminants. (ii) Due to the quick reaction of our reagents and the sample within 10 minutes, the overall processing time is significantly reduced. (iii) ExoPRISMTM reagents can be easily removed from the enriched EVs and allows elution by the buffer solution of your choice. (iv) As a result, ExoPRISMTM has no effect on downstream analysis or applications.
The kits include two types of reagents: Reagent B and Reagent C, which are optimized for CCS and urine samples, respectively. The Reagent B is an EV precipitation solution and the Reagent C is a washing solution.
List of Components

"Reagent B and C should be stored at room temperature. If there are precipitates in Reagent B, please warm it at 37 ℃ in the stirring incubator until the precipitates disappear with the naked eye, and cool it down at room temperature before use.

Usage of Reagents

The volume of reagents needed was calculated based on 5 ml of sample. Therefore, additional adjustment of the reagents should be required based on the sample volume.

The reagent volumes are calculated based on 5 mL of CCS or urine. The mixing ratio between sample and Reagent B should be kept at 1:1.Depending on the variety of biological samples, you can adjust the volume of Reagent C.