Exo-spin™ technology combines precipitation and size exclusion chromatography (SEC), making it superior to other techniques that rely solely on one method. Using only precipitation for exosome isolation will result in co-purification of large amounts of non-exosomal proteins and other material as well as carryover of the precipitant. SEC is reliable for exosome isolation, but a precipitation step is needed to concentrate your sample prior to SEC isolation. If there is no precipitation, a lower exosome recovery will be observed. We provide a simple two-step protocol which allows you to purify your sample in less than 2 hours with consistent and reliable results!

However, the EX04 Exo-spin™ kit only contains packed and equilibrated ready-to-use SEC columns as the precipitation step might not be recommended for some applications (e.g. mass spectrometry) or not needed for 1 ml of blood sample.

The EX04 Exo-spin™ kit has been developed to process up to 500 ml of cell culture medium, saliva, urine and 1 ml of blood samples (plasma and sera).

Which Exo-spin™ kit shall I choose for my sample volume?

* For cerebrospinal fluid (CSF) and human breast milk samples, validated protocols are available for EX01 only. The protocols are provided in the user guides.

What is size exclusion chromatography (SEC)?

SEC is a method to separate particles in solution based on their size. The chromatography column is packed with stable polymeric beads, creating a porous matrix. When the solution containing exosomes is added to the SEC column, the smaller particles will be trapped in the pores, while larger molecules do not enter the pores and elute first. As a result, different elution fractions will contain molecules of different sizes; first the large particles, followed by the smaller particles.

This technique is ideal to achieve highly purified samples in which exosomes are separated from other non-EV components, and it is compatible with both low and high initial sample volumes.

Exo-spin™ column and resin pore size

The column bed volume is 10 ml, allowing for 1 ml of volume to be loaded on top of the column.

The pore size of the resin is approximatively 30 nm to attain a highly pure exosome elution. All other molecules (e.g. proteins, lipids) which are smaller than 30 nm will enter into the pores and remain trapped in the column.

Highest recovery and purity

As mentioned above, essentially all proteins and lipids will be retained in the column and will elute later than the exosomes, ensuring a highly pure sample ready for your downstream application.

Reproducibility

All our columns are manufactured in our laboratory to ensure a high reproducibility between each lot. As proof of reproducibility and batch-to-batch consistency, a large number of peer-reviewed scientific papers have been published describing the use of Exo-spin™.

Storage

Upon receipt, store purification columns and Exo-spin™ Buffer at 4°C. All other components should be stored at room temperature (15°C – 25°C).

Frequently Asked Questions (FAQs)

For any additional questions, please refer to FAQs document below.

Start today! Select our starter pack

We designed the ideal starter pack to guide your exosome research. The starter pack includes the exosome purification kit of your choice, exosome validated antibodies, and NTA profiling analysis. The complete details can be found in the product page here.

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EX04 Exo-spin™ midi columns comparative data

Comparison of total exosome recovery from 10 ml conditioned cell culture medium. The total particle yield obtained with Exo-spin™ buffer + Exo-spin™ midi columns is greater than the yield obtained using the membrane affinity method. Notably, Exo-spin™ buffer is much more efficient than generic PEG 6000 when used prior to Exo-spin™ midi column exosome isolation.

Data determined by nanoparticle tracking analysis (NTA). Each curve represents the average of 3 technical replicate measurements for each exosome isolation method triplicate experiment. (PM = Precipitation Method). Figure taken and adapted from (Martins, TS et al., 2018).

Characterizing Exo-spin™ isolated exosomes using nanoparticle tracking analysis (NTA)

Exosome characterization with NTA. Exosomes have been isolated using the Exo-spin™ kit and analysis performed with the ZetaView® instrument.

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The EX04 Exo-spin™ kit is compatible with all downstream application and has been published in a large range of different applications.

RNA analysis:

• Santangelo L, Giurato G, Cicchini C, Montaldo C, Mancone C, Tarallo R, Battistelli C, Alonzi T, Weisz A and Tripodi M. (2016). The RNA-Binding Protein SYNCRIP Is a Component of the Hepatocyte Exosomal Machinery Controlling MicroRNA Sorting. Cell Reports 17(3):799-808.
• Brook AC, Jenkins RH, Clayton A, Kift-Morgan A, Baby AC, Shephard AP, Mariotti B, Cuff SM, Bazzoni F, Bowen T, Fraser DJ, and Eberl M. (2019). Neutrophil-derived miR-223 as local biomarker of bacterial peritonitis. Scientific Reports 9(1):10136.

Immunoassay:

• Salimu J, Webber J, Gurney M, Al-Taei S, Clayton A and Tabi Z. (2017). Dominant immunosuppression of dendritic cell function by prostate-cancer-derived exosomes. Journal of Extracellular Vesicles 6(1):1368823.

Protein assay:

• Welton JL, Webber JP, Botos LA, Jones M and Clayton A. (2015). Ready-made chromatography columns for extracellular vesicle isolation from plasma. Journal of Extracellular Vesicles 4:27269.

Application note:

• Comparison between precipitation and size exclusion chromatography vs. membrane affinity for exosome isolation.pdf

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[accordion title=”User guide”] EX04_Exo-spin_User_Guide[/accordion]

[accordion title=”FAQs”] FAQs[/accordion]
[accordion title=”MSDS/Tech Sheet”] EX03_and_EX04_MSDSDS.pdf[/accordion]
[accordion title=”Product Brochure”] CellGS exosome brochure.pdf[/accordion]
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