Proteomics Related Products

Exosomes Purification Column EVSecond


EVSecond/EVSecond L70

EVSecond is a size exclusion chromatography open column optimized for effective purification of exosomes. Highly-purified exosomes can be easily collected from serum, plasma, or cell culture supernatant. (EVSecond:Extracellular Vesicle Isolation by Size Exclusion Chromatography on Drip Column)


  • Simple gravity-flow handling without ultracentrifugation
  • EVSecond-purified exosomes possess efficient purity for comprehensive miRNA, proteome, and metabolome analysis
  • Exosomes are gently eluted in PBS without structural damage, allowing readministration experiments of collected exosomes to cells or animals
  • EVSecond L70 can used at room temperature and the preparation of column is now much easier


EVSecond L70
Column  Full Length
110 mm
75 mm
Packing Bed Length
70 mm
50 mm
In a refrigerator at 4 °C
In a refrigerator at 4 °C
Operation  Temperature
Room Temperature
at 4 °C
Column preparation
Room Temperature
Several minutes
at 4 °C
Several hours
Sample Volume
50 µL - 1,500 µL
50 µL - 700 µL
Wash after Blocking
PBS 1,500 µL 3 times
PBS 700 µL 6 times

Purification of Exosomes from Human Serum

A large amount of free proteins, metabolites, and nucleotides are involved in serum samples. Insufficient purification of exosomes often causes co-detection of non-exosomal components, leading to incorrect quantification results in omics studies. Exosomes were isolated from 100 µL, 200 µL, 300 µL, or 700 µL of human serum using EVSecond method. Exosomes were clearly separated from serum free proteins such as albumin or immunoglobulins.


100 µL/ fraction
Red line: CD9 - CD9 exosome sandwich ELISA (detecting exosomes)
Blue line: Bradford assay (detecting serum free proteins)

Operation flow

Gravity-flow is applied to each step.
1. Remove the top and bottom caps from the column and discharge the preservative solution from the column.
2. Filtrate the FBS through a 0.22 µm syringe filter. Then, load 700 µL of the filtrated FBS to the column.
3. Load 700 µL of PBS, 6 times to the column to wash.
4. Gently load 50 µL - 700 µL of serum sample which was prepared at the “sample preparation of serum sample” to the column.
5. Flow proper amount of PBS and collect exosomes eluent fraction.

Note) In first analysis, check the exosome elution position with anti-CD9-anti CD63 sandwich ELISA or anti-CD9 Western blotting. When the same sample is used for the second and subsequent rounds, it is not necessary to check the elution position.

Advantages over traditional procedures

  • Much higher-purity exosomes can be obtained compared to ultracentrifugation or polymer precipitation methods.
  • Unlike immuno-affinity purification using anti-tetraspanin antibodies, whole exosomes can be collected regardless of surface antigen profiles.

EVSecond L70RefrigerateENG10pcs5010-21395

GL-SPE EXO Fraction Rack

Open column rack optimized for EVSecond. It helps smooth column handling and fractionation.

Dimensions: 300(W) × 300(D) × 150(H) mm

  • Maximum 8 collection tubes(1.5 mL or 2 mL) can be set. The collection tube can be switched by pulling the lever.
  • Maximum 6 columns can be installed.
  • By adjust the height, it can use with both EVSecond and EVSecond L70.

GL-SPE EXO Fraction Rack1/pk5010-50450