Spin column 방식을 이용하여 serum, plasma와 cell culture supernatant에서 exosome 및 extracellular vesicle을 빠르고 쉽게 분리할 수 있습니다.
Intact vesicles are eluted from the exoEasy membrane with higher purity compared with ultracentrifugation.
Scanning electron microscopy (SEM; 20,000x magnification) was performed on a solubilized pellet from ultracentrifugation of pre-filtered (0.8 µm) plasma compared to an exoEasy eluate. Both preparations contain vesicle-shaped structures with an expected size range from 50–200 nm (white arrows; scale bar 200 nm). The preparation from ultracentrifugation also includes many smaller, unidentified structures that do not match the expected shape and size of extracellular vesicles.
Extracellular vesicles isolated using the exoEasy Kit are taken up efficiently by target cells (customer data).
HEK 293T cells were grown in vesicle-free DMEM for 66 h. Extracellular vesicles (EVs) were isolated using the exoEasy Kit from 15 ml cell culture supernatant pre-filtered using a 0.8 µm filter. Particle concentration was determined using the Nanosight instrument. Isolated EVs were labeled using BODIPY TR Ceramide for 20 min at 37°C. Unincorporated dye was removed by size exclusion chromatography (SEC). As a negative control, an equivalent amount of free dye in PBS was subjected to the same SEC cleanup. To test the uptake of EVs into target cells, approximately 1.5 x 109 labeled particles (representing about 1% of the total eluate from an exoEasy preparation started with 15 ml medium) were incubated with 2 x 104 HEK 293T cells in 200 µl DMEM for 1 h at 37°C. After washing, cells were stained with DAPI to visualize nuclei (bottom row) versus EVs taken up by cells (top row). Left column: negative control, right column: labeled EVs.
|exoEasy Maxi Kit||76064||20|
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