ExoPureTM Reagent (Overall Exosome Isolation, biological fluids)

Usage: RUO Cat: M1001-5, -10, -20; 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.

ExoPureTM is a fast and convenient one-step method of exosome isolation from biofluids (i.e. plasma, serum and urine) and from cell culture media.

Isolation with ExoPureTM is based on chemical precipitation. Samples are incubated with ExoPureTM solution on ice, so that exosomes will precipitate following centrifugation.

The obtained pellet can be resuspended in 1X PBS or deionized water. The protocol is user friendly, timesaving (around 1 hour), and does not require capital laboratory equipment.

usage

Application

  • Single step isolation of exosomes from plasma and serum
  • ExoPureTM is able to isolate the overall exosome population from small volumes of sample (as low as 100 μl of plasma)
  • Isolated exosomes are suitable for a wide range of analyses, such as nano tracking analysis (NTA), protein profiling by using different techniques (western blotting, ELISA, FACS), nucleic acid extraction and profiling of mRNA or small RNA markers
  • Easy to store and ships at 4°C

Sample Type

  • Human biological fluids including Plasma and Serum.

Kit Contents

Components M1001-5 (5mL) M1001-10 (10mL) M1001-20 (20mL)
ExoPureTM reagent (plasma/serum) 1 bottle(5 ml, 180 reactions for plasma, 80 reactions for serum) 1 bottle (10 ml, 350 reactions for plasma, 160 reactions for serum) 1 bottle (20 ml, 700 reactions for plasma, 320 reactions for serum)

ExoPureTM (Exosome Isolation from human plasma and serum) Assay Protocol: Volume suggested

Fluid Minimum volume required Volume suggested
Plasma 100 μl 100 μl - 250 μl
Serum 200 μl 250 μl - 500 μl
  1. Plasma and Serum samples preparation: Prepare plasma and serum samples by 3 centrifugation steps to eliminate red blood cells and cellular debris.
    1. 10 min at 300g
    2. 20 min at 1200g
    3. 30 min at 10,000g
  2. Exosome isolation:
    1. Add ExoPureTM solution to your sample in ratio 1/4 (i.e. 100 μl of plasma + 25 μl of ExoPureTM).
    2. Mix well by pipetting and inverting tube.
  3. Incubate on ice for 1 hr.
  4. Centrifuge 20 min at 10,000g (centrifuge can be performed at 4°C or at RT).
  5. Discard the supernatant.
  6. Centrifuge for 2 min at 1500g to eliminate entirely the supernatant.
  7. Resuspend the pellet in 100 μl * of 1X PBS.
  8. Resuspended exosomes can be used for analysis or stored at -20°C. * Volume of resuspension can be defined by the user on the base of downstream analysis.