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ExoIntact™ Exosome isolation kit

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Affinity selection by the peptide realizes higher purity than that in conventional methods such as ultracentrifugation orpolymer-precipitation that entangles unignorable amount of impurities.


Features

  • Extraction of highly pure EVs
  • Intact EVs are harvested thanks to the mild elution condition
  • Applicable to various downstream experiments

Outline of procedure

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Comparison experiment with conventional methods

Comparsion performance with others

EVs were isolated from serum. Then, isolation kits were compared by relative template ratio from Ct value of qPCR and protein concentration by BCA assay.

Method [1]Ralative abundance of EVs (Ct Value)#1 Protein Concentration (µg/mL) Purity([1]/[2]) Relative purity Required time scale(h)#2
Exolntact 1.00
(32.52)
45 2.2E-02 10491 2.0
Ultracentrifugation 0.25
(34.50)
15018 1.7E-05 8 2.5
Polymyer Precipitation 0.09
(35.93)
44815 2.1E-06 1#3 1.5
Figure
  • #1: Target sequence: hsa-miR-142-3p
  • #2: Starting from a pretreated sample
  • #3: Relative purity was calculated by relative template ratio over protein concentration and normalized so that polymer method is 1.

ExoIntact shows higher miRNA concentration and less protein concentration than conventional methods, which indicates that ExoIntact can collect more EVs with less protein impurities as calculated in Relative purity.

qNano: Particle Distribution

EV sample was prepared from one test of ExoIntact, exchanged the buffer to PBS by using centrifugal concentrator.

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TEM image

EV sample was prepared from one test of ExoIntact without further operations and sent it to Hanaichi Ultrastructure Research Institute to observe a shape of particles. A spherical particle was observed. Its diameter was about 75 nm and 120 nm. This result supports that ExoIntact successfully isolates EVs without damage.

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Kit contents

ExoIntact™ Exosome isolation kit (10 tests)

  • Streptavidin Magnetic Beads 200 μL x 1
  • Biotin-labeled EX peptide 200 μL x 1
  • Binding Buffer  2 mL x 1
  • BW Buffer 45 mL x 1
  • Elution Buffer 1 mL x 1
  • 1.5 mL Tubes 30 pcs/bag x 1

Sample preparation

Cell culture medium

  1. Culture the cells under appropriate conditions*1.
  2. Collect the cell culture medium.
  3. Centrifuge the medium for 5 minutes at 300 x g, 4℃ to remove cells and debris.
  4. Transfer the supernatant into a new centrifuge tube.
  5. Centrifuge the supernatant for 30 minutes at 10,000 x g, 4℃ to remove large EVs.
  6. Transfer the supernatant into a new centrifuge tube.
  7. Transfer 1.0 mL of the supernatant collected at step 6) into a 1.5 mL Tube and add 100 µL of Binding Buffer. (Please adjust the amount of Binding Buffer to be added depending on the amount of sample.)

*1: Fetal bovine serum (FBS) regularly contains bovine derived EVs. Please use EVs-depleted FBS if necessary.

Serum

Note: MicroSpin S-400 HR Columns is used in the following steps.

  1. Vortex the resin of MicroSpin S-400 HR Columns, and incubate it for 1 minute.
  2. Remove the cap, place the column into the provided collection tube, and centrifuge it for 1 minute at 735 x g, 4℃ to remove the excess storage buffer.
  3. Transfer the column to a new 1.5 mL Tube and add 100 µL of serum to the column carefully.
  4. Centrifuge it for 2 minutes at 735 x g, 4℃.
  5. Add 400 µL of BW Buffer to 100 µL of the collected eluate.

Download

ExoIntact Flyerpdf(PDF 713 KB)

ExoIntact Instruction pdf(PDF 341 KB)

Ordering Information

Product Name Storage Catalog Number PKG Size
ExoIntact™ Exosome isolation kit (10 tests) 4℃ S.051010 10 tests

Manufacturer : Siliconbio inc