GL-SPE EXO Fraction Rack

GL-SPE EXO Fraction Rack

Cat.No. FNK-501050450
Size 1 set

Description

Special Rack : GL-EXO Fraction Rack

• 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

Exosome purification column by size exclusion chromatography EVSecond

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.

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) 

(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.
 

 

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.

 

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