What experiments is Exosupur ® exosome purification kit-cell supernatant, etc. used for?

Exosupur ® exosome purification kit-cell supernatant, etc. is suitable for exosupur ® exosome purification kit is a purification kit specially used to extract small extracellular vesicles sEV from non-complex samples such as cell-culture supernatant, urine, bacterial culture supernatant, etc. The kit uses the principle of size exclusion chromatography to separate exosomes based on the molecular size of the components in the separated sample. This kit is rapid, efficient, simple to operate, and requires low equipment. It can complete the isolation and purification of exosomes in about 30-60 minutes from sample loading to fraction collection. The extracted exosome products have stable and reliable quality and low protein contamination, and can be applicable to downstream biological experiments such as protein extraction analysis, RNA extraction analysis, and cell function testing. Provide a systematic solution for the isolation of sEV from simple biological samples with small to medium-sized to large-volume cell supernatants.

Which product category does Exosupur ® exosome purification kit-cell supernatant, etc. belong to?

Exosupur ® exosome purification kit-cell supernatant, etc. belongs to the Exosome Purification Kits category and can be evaluated together with related exosome reagents for culture, isolation, purification, engineering, characterization, or tracing workflows.

Are specifications or documents available for Exosupur ® exosome purification kit-cell supernatant, etc.?

Yes. The page provides ES911.pdf, ES914.pdf, ES933.pdf, ES910.pdf. Detailed specifications should be confirmed with the page or official documents.

What should be confirmed before using Exosupur ® exosome purification kit-cell supernatant, etc.?

Users should confirm sample type, experimental goal, starting volume or particle concentration, downstream assays, and whether supporting purification, characterization, or tracing workflows are required.

Exosupur ® exosome purification kit-cell supernatant, etc.

Overview Regarding exosome purification methods, each method has advantages and disadvantages when performing exosome isolation according to different principles in the Minimum Experimental Requirements of the International Society for Extracellular Vesicles…

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Exosome Purification Kits
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Overview Regarding exosome purification methods, each method has advantages and disadvantages when performing exosome isolation according to different principles in the Minimum Experimental Requirements of the International Society for Extracellular Vesicles…

Overview

Separation method | Advantages and Disadvantages: Ultracentrifugation (DC) | Common exosome isolation methods; But first, special large-scale ultracentrifugation equipment is required; Secondly, in order to increase the yield, longer centrifugation time or greater centrifugal force is required, but this will inevitably increase the content of impurity proteins (usually proteins with a density similar to or higher than that of EVs) in the product; Thirdly, DC may cause aggregation or damage of EVs. In addition, the DC yield reported in the literature ranges from 2 to 80%. It is conceivable that the biochemical composition of the products obtained under different centrifugation conditions is difficult to maintain consistent and the stability is poor; Moreover, the ultracentrifuge operation is complex (the whole process takes about 3-9 hours) and the throughput is limited. It is only suitable for scientific research purposes and cannot meet the needs of clinical applications. / Density gradient centrifugation (DGC) | It can effectively remove impurity proteins in samples and is especially suitable for applications that require high product purity. However, this method is very complex to operate (the whole process ranges from 16 to 90 hours), the yield is low (about 10%), and the throughput is severely limited, making it only suitable for scientific research purposes. / Immunocapture (IC) | The product has high purity; However, it relies on antigen expression, and there is a preference for subpopulations; On the other hand, magnetic bead antibodies are expensive and are not suitable for processing large-volume samples; Finally, when using antibody magnetic beads to process samples such as plasma, the official recommendation of the kit is to utilize an SEC kit to pre-purify the sample. The disguised form demonstrates that the immune capture method is not ideal for direct processing of complex samples. / Polymer precipitation (PEG) | The operation is simple and the cost is low, the product EVs can be concentrated up to 50 times, and there is almost no limit on the sample volume. However, the selectivity is low and the product contains a large number of PEG molecules and impurity proteins. It is not recommended to consider this separation method for precious clinical samples.

Molecular size exclusion chromatography SEC stands out among many separation methods. It stands out due to its excellent performance and has won the favor of experts in the field of EVs. Its research proportion in the EVs special issue JEV is increasing day by day. The Exosupur ® kit, a patented product for the isolation and purification of exosomes based on the SEC principle developed by Echo biotech, has been tested by experts in the market and EVs field since its launch, and has received unanimous praise for its ease of operation, performance, stability and cost-effectiveness.

Overview

Regarding exosome purification methods, each method has advantages and disadvantages when performing exosome isolation according to different principles in the Minimum Experimental Requirements of the International Society for Extracellular Vesicles (ISEV) (MISEV2023): Separation method Advantages and Disadvantages Ultracentrifugation (DC) L Common exosome isolation methods; L But first, a special large-scale ultracentrifugation instrument is required; L Secondly, in order to increase the yield, longer centrifugation time or greater centrifugal force is required, but this will inevitably increase the content of impurity proteins (usually proteins with a density similar to or higher than that of EVs) in the product; L Thirdly, DC may cause the agglomeration or breakage of EVs. In addition, the DC yield reported in the literature is between 2-80%. It is conceivable that the biochemical composition of the products obtained under different centrifugation conditions is difficult to maintain consistent and the stability is poor; L Moreover, ultracentrifuges are complex to operate (the whole process takes about 3-9 hours) and have limited throughput. They are only suitable for scientific research purposes and cannot meet the needs of clinical applications. Density gradient centrifugation (DGC) L It can effectively remove impurity proteins in samples and is especially suitable for applications that require high product purity. L However, this method is very complex to operate (the whole process ranges from 16 to 90 hours), the yield is low (about 10%), and the throughput is severely limited, making it only suitable for scientific research purposes. Immunocapture (IC) L Product purity is high; L However, depending on antigen expression, there is a preference for subgroups; L On the other hand, magnetic bead antibodies are expensive and not suitable for processing large volumes of samples; L Finally, when using antibody magnetic beads to process samples such as plasma, the official recommendation of the kit is to utilize an SEC kit to pre-purify the sample. This demonstrates that the immunocapture method is not ideal for direct processing of complex samples. Polymer precipitation (PEG) L The operation is simple and the cost is low, the product EVs can be concentrated up to 50 times, and there is almost no limit on the sample volume. L However, the selectivity is low and the product contains a large number of PEG molecules and impurity proteins. It is not recommended to consider this separation method for precious clinical samples. Ultrafiltration (UF) L can effectively concentrate EVs by more than 200 times. L Generally only used as a means of concentration in conjunction with other methods. Size exclusion chromatography SECC) L There are no physical and chemical factors such as shearing force, retaining the integrity and biological activity of EVs without any impact on EVs. The particle size is uniform, the repeatability is robust, and the operation is simple and rapid, and it can effectively remove impurity particles with smaller particle sizes in the sample; L However, the product concentration is low, so it generally needs to be concentrated with ultrafiltration; L Moreover, the loading volume is limited by the column bed volume, so IF it is a large volume sample, it can be concentrated first and then loaded. Molecular size exclusion chromatography SEC stands out among many separation methods. It stands out due to its excellent performance and has won the favor of experts in the field of EVs. Its research proportion in the EVs special issue JEV is increasing day by day. The Exosupur ® kit, a patented product for the isolation and purification of exosomes based on the SEC principle developed by Echo biotech, has been tested by experts in the market and EVs field since its launch, and has received unanimous praise for its ease of operation, performance, stability and cost-effectiveness.

Specifications

• Exosupur ® sEV separation kit-10ml exclusion column Exosupur ® sEV separation kit-10ml exclusion column Specifications: ES910【10ml*1pcs】 1) Single loading 10ml*5 repetitions =50ml Application: Suitable for non-complex large-volume samples such as cell-culture supernatant and urine. Used with the ES8500 cell-culture supernatant high concentration kit, the total processing volume is up to 1.5L-2.5L • Exosupur ® sEV separation kit-1ml exclusion column Exosupur ® sEV separation kit-1ml exclusion column Specifications: ES911 & ES914 1) ES911 [1ml*1pcs], single loading 1ml*5 repetitions =5ml 2) ES914 [1ml*4pcs], single loading 1ml*4 tubes*5 times =20ml Application: Suitable for sEV purification of small volume samples (5-20ml) after concentration of non-complex cell supernatants, urine, etc. • Exosupur ® sEV separation kit-3ml exclusion column Exosupur ® sEV separation kit-3ml exclusion column Specifications: ES933 1) ES933 [3ml*3pcs], single loading 3ml*3 tubes*5 repetitions =45ml Application: Suitable for sEV purification of samples within 45ml after concentration of non-complex cell-culture supernatant, urine, etc. It is recommended to purchase exclusion column companions: brackets, adapters and other accessories - to increase operating throughput or facilitate liquid addition, link: Exosupur ® Exclusion column companions - brackets & adapter accessories - Echo biotech

Applications

• Compatible with different sample types such as cell-culture supernatant and urine; • can be applicable to high-throughput sequencing, mass spectrometry and other omics research on different samples, as well as downstream PCR, WB, WLISA verification, etc., with higher purity, less interference from impurities, and more accurate quantification; • It can be used to study the functional mechanisms of cells and animals without affecting the properties and activity of exosomes, or introducing aggregation and other impurities; • can be applicable to research such as re-purification of engineered exosomes after modification; • can be used as a control method in purification-related method development.

Advantages

• Easy to operate: no special equipment is required, and the four-step operation of column equilibration, sample loading, exosome collection, and exosome concentration only takes 30 -60 minutes from sample loading to completion of separation. The upgraded product is equipped with an adapter for column balancing or cleaning, making operation easier and freeing your hands; • Superior performance: up to 95% of cell-culture supernatant samples can be recovered, and the impurity protein removal rate is over 99%; • High stability: only exosomes in a fixed size range are recovered, and the product particle size range is narrow; the experiment is relatively less affected by fluctuations in external factors; the complete exosome morphology and function can be maintained without introducing other physical and chemical factors; • High cost-effectiveness: the exclusion column can be cleaned and regenerated and used repeatedly for 5 times, and the sample volume that can be processed is large; • Reliable quality: The Exosupur ® series of sEV purification kits are efficient and convenient. They have assisted customers in publishing 100+ articles. They have been inspected by industry experts and have been recognized as new technologies and new products by Beijing Municipality.

Performance

Characterization of TEM electron microscopy, WB and nanoFCM particle size of small extracellular vesicles sEV isolated from Exosupur ® cell-culture supernatant. For cell-culture supernatant samples, Exosupur ® can recover up to 95% of exosomes and has a removal rate of over 99% of impurity proteins. The lower left picture demonstrates the effect of separating 1 mL of concentrated cell-culture supernatant with a 1 ml exclusion column (ES914, ES911). The lower right picture demonstrates the effect of separating 3 mL of concentrated cell-culture supernatant with a 3 ml exclusion column (ES933). The second row below demonstrates the effect of 10 ml of concentrated cell-culture supernatant loaded on a 10 ml exclusion column. High-purity exosomes are more conducive to RNAomics, proteomics, cell function and in vivo experimental research. Protein K and RNase A double-enzyme digestion further confirmed that the RNA obtained by Exosupur ® was more derived from vesicles, while the detection value of free RNA outside the vesicles was lower. When conducting proteome research on exosomes isolated from Exosupur®, abundant vesicle-specific biomarker proteins can be detected, indicating that the exosomes are highly pure and can detect more proteins derived from exosomes; a relatively large number of proteins can be detected, which is conducive to differential analysis between different samples; there are many overlaps with published articles and databases, indicating that the data is reliable and can accurately study exosome proteins. Comparing different SEC separation methods: the fraction collected by the Exosupur ® kit has the highest fluorescence intensity (A) and the lowest protein concentration (B and C), indicating that Exosupur ® has the most effective recovery rate of EVs and the lowest free protein contamination.

FAQ

Q: can the Exosupur ® exosome purification kit be equipped with a similar plastic test tube rack for easy utilize? A: We have matching exclusion column brackets (ES914S and ES933S), which need to be purchased separately. Bracket link: Exosupur ® Exclusion Column Companion-Bracket & Adapter Accessories-Echo biotech Q: can the Exosupur ® exosome purification kit be equipped with a buffer for each multiple column volumes of PBS or cleaning solution flushing, so that it does not need to be added repeatedly? A: We have matching adapters (ES91A and ES93A), which need to be purchased separately. Adapter link: Exosupur ® Exclusion Column Companion-Stand & Adapter Accessories-Echo biotech Q: The adapter and the exclusion column cannot fit tightly, is there leakage? A: Adjust the direction of the adapter. The adapter should be inserted a little tighter. Normally, it will not affect the utilize. Q: What do I need to prepare for this kit to isolate exosomes? A: 1) Sterile PBS solution (must be filtered with a 0.22μm filter membrane. It is recommended to prepare it freshly or purchase membrane-passed PBS. PBS solutions stored in a 4°C refrigerator must be equilibrated to room temperature first); 2) 20% absolute ethanol (must be degassed under ultrasonic conditions for 20 minutes or left overnight after preparation); 3) Ultrafiltration tube (molecular weight cutoff 100kD); 4) Collection tubes (1.5mL EP tube and 15mL centrifuge tube); 5) Sample (needs to be equilibrated to room temperature before loading). Q: After collecting the samples, the color is still in the middle. Is there any problem? A: IF you follow the operating instructions, there will be no problem. Generally, the color lags because the color is composed of small molecules and will come out later, and the exosomes will flow out first, so the color cannot be used to judge the collection of exosomes. Q: Should I flush it directly with blocking solution after utilize, or flush it with 2 times the column volume of PBS and then flush it with 1 times the column volume of blocking solution? A: IF it is not going to be used this time, there is no need to wash it with blocking solution; IF it is not going to be used this time and needs to be saved for next time, then it should be rinsed with PBS and then rinsed with blocking solution this time. Q: How to store the isolated exosomes? A: The problem of exosome preservation has nothing to do with the isolation method, but is determined by its own characteristics. IF conditions permit, it is best to utilize fresh exosomes for experiments. Frozen exosomes are prone to agglomeration and precipitation. In actual research, exosome functional experiments often require a large amount of exosomes that have to be extracted and stored in batches and can be stored at -80°C; however, when doing biomarker research, such as RNAomics, follow-up experiments such as RNA extraction must be completed as soon as possible after exosomes are extracted. Q: can Exosupur® isolate exosomes derived from bacteria? A: Exosomes derived from bacterial culture supernatant can be isolated. The operation tutorial is as follows: ES914 Isolation of Helicobacter pylori Outer Membrane Vesicles OMV Citations: Wei S, Li Q: can Exosupur® isolate plant-derived exosomes? A: Yes, you can refer to: https://mp.weixin.qq.com/s/uZquvHdPzGLnaHpxe907Tg Q: can Exosupur® isolate milk-derived exosomes? A: Yes, you can refer to: https://mp.weixin.qq.com/s/-PiBtUqg2wOcozeclJhhUw

Manual Downloads

References

At present, the Exosupur® exclusion kit has been cited in 100+ SCI literatures, so you can purchase it with confidence. 1、Li X, Cui J, Ding Z, et al. Klebsiella pneumoniae-derived extracellular vesicles impair endothelial function by inhibiting SIRT1. Cell Commun Signal. 2025;23(1):21. IF=8.9 Literature download link: https://biosignaling.biomedcentral.com/articles/10.1186/s12964-024-02002-0 2、Wang C, Zhou X, Bu T, et al. Engineered extracellular vesicles as nanosponges for lysosomal degradation of PCSK9. Mol Ther. Published online November 26, 2024. doi:10.1016/j.ymthe.2024.11.034 IF=12.0 Literature download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABA5icrKY4 3、Liu L, Chen S, Liang S, Liang Z. The expression profile of brain-derived exosomal miRNAs reveals the key molecules responsible for spontaneous motor function recovery in a rat model with permanent middle cerebral artery occlusion. Mamm Genome. 2024;35(3):362-376. Literature download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABASeS5Dil 4、Chen R, Kang Z, Li W, et al. Extracellular vesicle surface display of αPD-L1 and αCD3 antibodies via engineered late domain-based scaffold to activate T-cell anti-tumor immunity. J Extracell Vesicles. 2024;13(7):e12490. IF=14.5 Document download link: https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jev2.12490

Documents

ES911.pdf

ES914.pdf

ES933.pdf

ES910.pdf

FAQ

What is Exosupur ® exosome purification kit-cell supernatant, etc.?: Exosupur ® exosome purification kit-cell supernatant, etc. is an exosome research reagent under the Exosome Purification Kits category. Overview Regarding exosome purification methods, each method has advantages and disadvantages when performing exosome isolation according to different principles in the Minimum Experimental Requirements of the International Society for Extracellular Vesicles…
What experiments is Exosupur ® exosome purification kit-cell supernatant, etc. used for?: Exosupur ® exosome purification kit-cell supernatant, etc. is suitable for exosupur ® exosome purification kit is a purification kit specially used to extract small extracellular vesicles sEV from non-complex samples such as cell-culture supernatant, urine, bacterial culture supernatant, etc. The kit uses the principle of size exclusion chromatography to separate exosomes based on the molecular size of the components in the separated sample. This kit is rapid, efficient, simple to operate, and requires low equipment. It can complete the isolation and purification of exosomes in about 30-60 minutes from sample loading to fraction collection. The extracted exosome products have stable and reliable quality and low protein contamination, and can be applicable to downstream biological experiments such as protein extraction analysis, RNA extraction analysis, and cell function testing. Provide a systematic solution for the isolation of sEV from simple biological samples with small to medium-sized to large-volume cell supernatants.
Which product category does Exosupur ® exosome purification kit-cell supernatant, etc. belong to?: Exosupur ® exosome purification kit-cell supernatant, etc. belongs to the Exosome Purification Kits category and can be evaluated together with related exosome reagents for culture, isolation, purification, engineering, characterization, or tracing workflows.
Are specifications or documents available for Exosupur ® exosome purification kit-cell supernatant, etc.?: Yes. The page provides ES911.pdf, ES914.pdf, ES933.pdf, ES910.pdf. Detailed specifications should be confirmed with the page or official documents.
What should be confirmed before using Exosupur ® exosome purification kit-cell supernatant, etc.?: Users should confirm sample type, experimental goal, starting volume or particle concentration, downstream assays, and whether supporting purification, characterization, or tracing workflows are required.