Cell Guidance Systems Exo-spin Mini-Columns Panduan Pengguna - Halaman 4

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Product Information

Exo-spin™ columns may be used for isolation of intact exosomes from any sample

smaller than 100 µl. This includes blood sera, but not blood plasma. Exo-spin™

columns may be used for purification of samples which have been purified by other

methods, such as precipitation and ultracentrifugation, in cases where additional

cleanup is required to remove proteins and smaller molecules. Exo-spin™ columns

are very effective for the clean-up of labeled exosome samples.

Exosomes isolated using Exo-spin™ columns may be used in a variety of

downstream applications including DNA and RNA studies as well as in functional in

vitro and in vivo exosome assays.

Plasma contains much higher protein levels than sera and can only be processed

directly using EX04 Exo-spin™ Midi columns. Also, a kit specifically for isolation of

exosomes from blood (EX02 Exo-spin™ blood) can be used for isolation of

exosomes from plasma.

General Information

Notes on Cell Culture

Fetal bovine serum (FBS) contains a large number of exosomes. Exosome-free FBS

should be used in cell culture experiments. Exosome-free FBS is available

commercially. Alternatively, we found that Vivaspin20 100,000 MWCO PES (GE

Healthcare) centrifugal concentrators or Millipore® UFC910024 Amicon® Ultra-15

Centrifugal

Filter

Concentrator

with

Ultracel®

100

Regenerated

Cellulose

Membrane, NMWL: 100,000 can be used to efficiently remove exosomes from FBS,

which should be previously diluted 50% in PBS.

The number of exosomes that are obtained from a cell culture sample will vary

depending on a variety of factors. These include the cell line, the length of time the

medium is exposed to the cells, and the total number of cells in culture. Cancer cell

lines may produce higher numbers of exosomes than non-transformed cell lines.

Note on collection of samples

A review in 2013 by Witwer et al (Journal of Extracellular Vesicles (2013) 2: 20360)

indicates that the way samples are collected and handled prior to purification can

have a significant impact on the quality of purified exosomes.