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)

Features

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

Specifications

	EVSecond L70	EVSecond
Column Full Length	110 mm	75 mm
Packing Bed Length	70 mm	50 mm
Storing	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)
Data provided by Dr. Koji Ueda. at JAPANESE FOUNDATION FOR CANCER RESEARCH.

Operation flow

Instruction Manual

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, 3 times to the column to wash.
4. Gently load 50 µL - 1500 µL of serum sample which was centrifuged (12,000 × g, 4℃, 15min).
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. Furthermore, check the serum free protein elution position with Bradford method. When the same sample use 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.

Description	Transportation	SDS	Qty.	Cat.No.
EVSecond L70	Refrigerate	ENG	10pcs	5010-21395
EVSecond	Refrigerate		10pcs	5010-21390 Discontinued
EVSecond	Refrigerate		25pcs	5010-21392 Discontinued

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.

Description	Qty.	Cat.No.
GL-SPE EXO Fraction Rack	1/pk	5010-50450

5010-50450