ExoPureTM Chromatography Columns

Usage: RUO Cat: K1246-5, -10; Size Exclusion Chromatography (SEC) Columns; Store at 4°C

Introduction

Exosomes are small endosome derived lipid nanoparticles (50-120 nm) actively secreted by exocytosis by most living cells

Exosome release occurs either constitutively or upon induction, under both normal and pathological conditions, in a dynamic, regulated and functionally relevant manner. Both the amount and molecular composition of the released exosomes depend on the state of a parent cell.

Exosomes have been isolated from diverse cell lines (hematopoietic cells, tumor lines, primary cultures, and virus infected cells) as well as from biological fluids in particular blood (e.g. serum and plasma from cancer patients) and other body fluids (broncho alveolar lavage fluid, pleural effusions, synovial fluid, urine, amniotic fluid, semen, saliva etc). Exosomes have pleiotropic physiological and pathological functions and an emerging role in diverse pathological conditions such as cancer, infectious and neurodegenerative diseases.

Size Exclusion Chromatography (SEC) is considered one of the best methods for isolating and purifying exosomes and extracellular vesicles (EVs) from different matrices. This technique is very efficient for separating EVs from the circulating proteins without affecting the original shape and functionality of the vesicles. ExoPureTM Chromatography Columns is a SEC column designed for isolating EVs in a fast and easy way.

usage

Application

  • For the isolation of highly pure exosomes from biofluids (plasma, serum, urine) and cell media
  • For EV purification of pre-isolated EVs from contaminants (500 µl up to 2 ml)
  • Isolated exosomes are suitable for multiple analysis including NTA, ELISA, FACS, WB, EM, MS, nucleic acid extraction etc
  • Easy and fast protocol (approximately 15 minutes)
  • Isolate exosomes from different samples types
  • Easy to store and ship at 4°C

ExoPureTM SEC Purification Protocol

Fluid Volume Amount
Plasma 0.5 ml up to 2 ml
Serum 0.5 ml up to 2 ml
Urine 0.5 ml up to 2 ml (concentrated to 10-fold)
Cell Media 0.5 ml up to 2 ml (concentrated to 10-fold)

Prepare the sample by centrifugation steps as suggested in the table below

|Fluid|Suggested|Optional| |-|-|-|-| |Plasma|10 min at 300g (save super); 20 min at 1200g (save super)|30 min at 10000g (to eliminate vesicles > 200 nm)| |Serum|10 min at 300g (save super); 20 min at 1200g (save super)|30 min at 10000g (to eliminate vesicles > 200 nm)| |Urine|10 min at 300g (save super); Concentrate 10-fold using concentrators|| |Cell Media|10 min at 300g (save super); 20 min at 1200g (save super) Concentrate 10-fold using concentrators|Centrifuge before using concentrators, 30 min at 10000g (to eliminate vesicles > 200 nm)|

Other biofluids which present a diluted population of EV can be concentrated 10-fold using concentrators. Viscous fluids (saliva) must be diluted in 1X PBS before proceeding to the EV isolation in SEC columns.