The exoEasy Kit improves upon traditional ultracentrifugation exosome or microvesicle isolation methods, yielding purified extracellular vesicles in just 25 minutes.
The exoEasy Maxi Kit uses a membrane-based affinity binding step to isolate exosomes and other EVs from serum and plasma or cell culture supernatant. The method does not distinguish EVs by size or cellular origin, and is not dependent on the presence of a particular epitope. Instead, it makes use of a generic, biochemical feature of vesicles to recover the entire spectrum of extracellular vesicles present in a sample (see figure “ Characterization of extracellular vesicles isolated using the exoEasy Maxi Kit by NTA
”). It is therefore essential to completely remove cells, cell fragments, apoptotic bodies, etc., by centrifugation or filtration of samples before starting the protocol.
The technology developed with Exosome Diagnostics, Inc., uses a spin column format and specialized buffers to purify exosomes from pre-filtered biological fluids; up to 4 ml using plasma or serum. For cell culture supernatants, processing of up to 32 ml sample (4 column loading steps) has been successfully tested. However, the concentration of vesicles in supernatants depends strongly on the cell type and culture conditions; therefore, we recommend starting with no more than 16 ml of supernatant for sample material that has not been tested with the kit previously. Higher sample volumes may result in reduced recovery of vesicles. The procedure is very fast, consistent and highly suited for functional analysis of exosomes and other extracellular vesicles. Particulate matter other than vesicles, such as larger protein complexes that are especially abundant in plasma and serum, is largely removed during the binding and wash steps.