EVSecond Exosome Purification

Product#: 501021392
$1,100.00
Availability:
Ships in 1-2 weeks

EVSecond Exosome Purification

Cat.No. 501021392
Size 25 pieces 

Description

Exosome purification column by size exclusion chromatography EVSecond
501021390.png

EVSecond is a specially developed size exclusion chromatography (SEC) column, for Exosome purification.
Exosome can be easily purified from serum, plasma and cell culture supernatant.
Special instrument, such as ultracentrifuge is not required.

Features

  • Easy procedure by free fall
    - Special instruments are not required.
  • Best for exhaustive miRNA analysis and high sensitivity proteome analysis
    - Free RNA or protien in sample can be efficiently removed.
  • Exosome can be applied to further application
    - Purified exosome do not have structural damage.
  • Special rack "GL-SPE EXO Fraction Rack" is available as an option
    - This special rack makes preparation and collection easy.
Example Data
501021390_fig1.png
Fig. 1 Purification of exosome from human serum
Each 100 μL of fraction are collected. By using EVSecond, exosome were purified from 100, 200, 300 or 700 μL human serum.

Exosome (Red) and serum proteins (Blue, e.g. albumin or IgG) are separated accurately.
Red : Exosome fraction site was confirmed by CD9 sandwich ELISA.
Blue : Serum proteins fraction site was confirmed by Bradford method.
Data is kindly provided by Dr. Ueda (University of Tokyo) 

specification
 

EVSecond L70

EVSecond

Column length

110 mm

75 mm

Filling length

70 mm

50 mm

Preservation

Refrigerated storage

Refrigerated storage

operation

room temperature

Low temperature room

Invert mixing

Room temperature
several minutes

Low temperature room
several hours

Sample volume

50 to 1,500 μL

50 to 700 μL

Washing after blocking

PBS 1,500 μL × 3 times

PBS 700 μL × 6 times

 
Procedure
(A) Preparation of Column (To be executed in 4 degree condition)
Mix filler well by inverting column until filler is mixed evenly and completely. 
* Mix the column on the shaker overnight gently in cold room is recommended.
* Filler clumps may cause incorrect fraction. Check no clumps are found before use.

(B) Purification (To be executed in 4 degree condition)

  1. Remove two caps (top and bottom) and discard storage buffer.
  2. Block the filler by 700 μL of FBS (filtrated by 0.22 μm)
  3. Wash 6 times by 700 μL of PBS
  4. Apply 50 to 700 μL of sample. Flow all sample completely.
  5. Apply 100 μL of PBS twice
  6. Apply 750 to 1000 μL of PBS (total added volume from 4 to 6 should be 1000 μL). Flow all volume and discard it.
  7. Prepare collection tube, and collect 12 fractions by 100 μL of PBS.
Note
First, confirm the exosome elution position by anti-CD9-anti-CD63 sandwich ELISA, anti-CD9 western blotting, etc. If you use the same sample from the second time on, you do not need to check the elution position.
 
501021390_fig2.jpg
 
Attention
* Initially, elution position of exosome and serum proteins should be checked by ELISA (CD9 ELISA, for exosome) or Bradford method (for serum proteins).

* Samples which contain objects and may stuck in the column should be pretreated by following protocol.

  1. Centrifuge sample by 12000 x g, for 15 minutes in 4℃.
  2. Collect sample from middle layer (50 to 700 μL). Do not take precipitation at the bottom or lipids (upper layer). 
Comparison with conventional method
  • Exosomes of higher purity can be isolated compared to ultracentrifugation and polymer precipitation.
  • Unlike antibody isolation methods that capture specific exosome surface antigens, it allows for less biased exosome recovery independent of surface antigen profiles.
Note
Refrigerated is a refrigerated shipment. Separately, we will charge you a refrigerated packing fee of 1,000 yen.
 

5010-21390.jpg

5010-21390.jpg

501021390_fig2png

501021390

501021392

501050450

 
Special Rack : GL-EXO Fraction Rack
501021390_fig2png
  • Rack can hold up to 8 collection columns (1.5 mL or 2 mL).
  • Collection columns can be shifted by manipulating a lever.
  • Up to six sets of columns can be set.
  • Size : 300(W)×300(D)×150(H) mm

Logo_GLSciences.png

Satisfaction
Quality Rating
Value Rating
Style Rating
X