Are specifications or documents available for Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples?

Yes. The page provides ES9P11e说明书.pdf, ES9P14e说明书.pdf, ES9P021e.pdf, ES9P024e.pdf. Detailed specifications should be confirmed with the page or official documents.

Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples

Overview In the minimum experimental requirements (MISEV2023) recommended by the International Society for Extracellular Vesicles (ISEV), the recommendations for isolating EVs based on commercial kits are as follows: Kits based on the principle of precipitati…

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Exosome Purification Kits
Products
Overview In the minimum experimental requirements (MISEV2023) recommended by the International Society for Extracellular Vesicles (ISEV), the recommendations for isolating EVs based on commercial kits are as follows: Kits based on the principle of precipitati…

Overview

Kits based on the principle of precipitation will concentrate all EP in the mixture, even many free proteins, resulting in highly impure preparations, especially from complex NVEP-rich sources such as plasma and serum. The utilize of such kits is strongly not recommended unless solely for volume reduction (concentration).

Principle of molecular size exclusion chromatography SEC: Biological fluid samples are added to an exclusion column containing SEC packing materials with fixed pores. Molecules with smaller particle sizes such as proteins and lipids can enter the fixed pores of the packing material, while molecules or particles with larger particle sizes cannot enter the pores and can only enter the gaps of the packing material. Therefore, with the addition of buffer, vesicles with larger particle sizes can only enter the interstitial space of the material and are eluted in the previous fractions due to their rapid flow rate, while small molecule impurities require a longer elution time to flow out, thus achieving the separation of EVs.

Overview

In the minimum experimental requirements (MISEV2023) recommended by the International Society for Extracellular Vesicles (ISEV), the recommendations for isolating EVs based on commercial kits are as follows: Kits based on the principle of precipitation will concentrate all EP in the mixture, even many free proteins, resulting in highly impure preparations, especially from complex NVEP-rich sources such as plasma and serum. The utilize of such kits is strongly not recommended unless solely for volume reduction (concentration). Principle of molecular size exclusion chromatography SEC: Biological fluid samples are added to an exclusion column containing SEC packing materials with fixed pores. Molecules with smaller particle sizes such as proteins and lipids can enter the fixed pores of the packing material, while molecules or particles with larger particle sizes cannot enter the pores and can only enter the gaps of the packing material. Therefore, with the addition of buffer, vesicles with larger particle sizes can only enter the interstitial space of the material and are eluted in the previous fractions due to their rapid flow rate, while small molecule impurities require a longer elution time to flow out, thus achieving the separation of EVs. Molecular size exclusion chromatography SEC has higher comprehensive performance among many separation methods, and its frequency of utilize is second only to ultraionization. Its research proportion in the EVs special issue JEV is increasing day by day. Exosupur ® Plasma/Serum sEV Purification kit is developed based on the SEC method and is a dedicated isolation and purification kit for sEV exosomes from complex biofluid samples such as plasma/serum.

Specifications

1) Exosupur ® sEV separation kit-1ml plasma/serum exclusion column Specifications: ES9P11e & ES9P14e 1) 1ml*1pcs, load 1ml*5 repetitions = 5ml, 5 samples 2) 1ml*4pcs, load 1ml*4 pieces*5 times = 20ml, 20 samples 2) Exosupur ® sEV separation kit-micro plasma/clearance exclusion column Specifications: ES9P021e & ES9P024e 1) 0.2ml*1pcs, load 0.2ml*5 repetitions = 1ml, 5 micro samples 2) 0.2ml*4pcs, load 0.2ml*4 tubes*5 repetitions = 4ml, 20 micro samples 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

• 1ml and microexclusion columns are suitable for complex biological samples such as normal or trace plasma/serum respectively; • The isolated sEVs derived from complex biological samples have higher purity and less interference from impurities. They are suitable for omics research such as high-throughput sequencing and mass spectrometry, as well as downstream expression quantitative research such as PCR, WB, and WLISA verification, and the quantification is more accurate; • This kit is used to isolate sEVs derived from complex biological samples. The method is gentle and simple, does not affect the properties and activity of exosomes, does not introduce aggregation and other impurities, and can be applicable to functional mechanism research experiments in cells and animals; • This kit can be used as a control method during method development related to sEV purification from complex samples.

Advantages

• Easy to operate: no special equipment is required, just 4 steps of column equilibration, sample loading, exosome collection, and exosome concentration. It only takes 30-60 minutes from sample loading to completion of separation; • Superior performance: For normal plasma samples, the Exosupur ® 1ml plasma/clearance exclusion column can recover up to 95% of fluorescent liposomes, and the impurity protein removal rate is as high as 99.5%; the trace plasma/clearance exclusion column can recover up to 94% of nanoparticles, and the average impurity protein removal rate is 98%; • 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; other physical (such as ultra-high-speed centrifugal force of ultrasonic) and chemical factors (such as PEG particles in the precipitation method) are not introduced, and the complete exosome morphology and function can be maintained; • High cost performance: By cleaning and regenerating the exclusion column, it can be reused 5 times, that is, a single exclusion column can process 5 samples; • Reliable quality: The Exosupur ® series of sEV purification kits are efficient and convenient. They have helped customers publish 100+ articles, including many high-quality documents with an IF of 10 or above. They have been inspected by industry experts and have been recognized as new technologies and new products in Beijing.

Performance

1. Performance data of Exosupur ® plasma exclusion column for separating plasma exosomes Characterization of TEM electron microscopy, WB and nanoFCM particle size of isolated plasma or trace plasma small extracellular vesicles sEV by Exosupur ®. The Exosupur ® 1ml plasma/clearance exclusion column can recover up to 95% of fluorescent liposomes from plasma samples, and the impurity protein removal rate is as high as 99.5%; the trace plasma/clearance exclusion column can recover up to 94% of nanoparticles, and the average impurity protein removal rate is 98%; The lower left picture demonstrates the effect of 1ml plasma/clearance exclusion column (ES9P14e, ES9P11e) in separating 1mL plasma sEV, and the lower right picture demonstrates the effect of trace plasma/clearance exclusion column (ES9P021e, ES9P024e) in separating 0.2mL trace plasma sEV. In the second row of pictures below, it can be seen from Western Blot that there are fewer major miRNA-binding proteins and fewer impurity proteins in plasma. High-purity exosomes are more conducive to RNAomics, proteomics, cell function and in vivo experimental research. 2. Exosupur ® plasma exclusion column and other principle plasma exosome isolation kits---competitive product analysis Using the Exosupur ® plasma exclusion column and two commercially available plasma exosome isolation kits with different principles (membrane affinity method kit-Q and precipitation method kit-S), as well as classic ultra-separation, various indicators of separated plasma exosomes were detected. The results are as follows: The isolated plasma sEV was loaded with proteins such as WB to detect the main contaminating proteins in the blood: albumin HSA, RNA-binding protein Ago2, and lipoprotein ApoB. The results showed that the exosomes separated by Exosupur ® had lower impurity protein levels. (The relative purity of plasma exosomes separated by kit-S based on the precipitation method is worse) The particle size distribution and particle number of sEV isolated from 1ml plasma were measured by NanoFCM. The results showed that the size distribution of plasma EV isolated by Exosupur ® was relatively narrow and consisted of small vesicles, with a moderate particle number yield. (The size of plasma exosomes separated by kit-S based on the precipitation method is relatively larger and has the highest yield) The purity of exosomes was evaluated by the ratio of the number of isolated plasma sEV particles to the protein content. The results showed that the exosomes isolated by Exosupur ® had higher purity and lower impurity protein content. Data from J Transl Med. 2021 Mar 12;19(1):104. For low-expression long RNA, Exosupur ® can obtain higher abundance of long RNA, which is more conducive to mRNA + LncRNA research. For miRNA, due to the higher yield and higher purity of exosomes enriched by Exosupur ®, the quantitative value of miRNA enriched in sEV is higher, while the quantitative value of non-enriched miRNA is very low, allowing more accurate analysis of sEV-derived miRNA. 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. 3. Exosupur ® Plasma Exclusion Column and Plasma exosome Exclusion Purification kit with the same SEC principle---Competitive Product Analysis The fraction collected by the Exosupur ® kit has the highest particle fluorescence intensity (i.e. particle concentration), the lowest protein concentration (A and B), and the highest particle/protein purity index (C), indicating that Exosupur ® has the most effective EVs recovery rate and the lowest free protein contamination, and the separated EVs have the highest purity. Note: The protein content is determined by the BCA method, and the number of particles is determined by detecting the A480 value. Panels D and E: Electron microscopy TEM and WB characterization results of exosomes derived from plasma, urine and cell-culture supernatant isolated using ES9P11e.

FAQ

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 need to 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 (it needs to be equilibrated to room temperature before loading, and the single loading volume of a single column is 1mL). Q: Which kit should be used IF the plasma sample is about 0.5ml? A: Directly dilute to 1ml and load onto Exosupur ® ES9P11e/ES9P14e 1ml plasma exclusion column Q: Does the 0.2ml sample loaded on the Exosupur ® Micro-Plasma/Clear Exclusion Column refer to the original plasma/serum? A: It refers to the sample that has been centrifuged at low speed and filtered through a 0.45 μm filter membrane, 0.2ml. Please note that approximately 100 μl will be lost by membrane filtration. IF only 0.2ml of plasma is applicable to membrane filtration, it is recommended that the customer utilize about 100 μl of PBS for a second pass through the membrane. 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 rinse directly with blocking solution after utilize, or rinse with 2 volumes of PBS and then rinse with 1 times the 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 isolated exosomes A: The issue of exosome storage 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.

Manual Downloads

References

• 1. Yin H, Xie J, Xing S, et al. Machine learning-based analysis identifies and validates serum exosomal proteomic signatures for the diagnosis of colorectal cancer. Cell Rep Med. 2024;5(8):101689. IF=10.6 Zone 1 1. Yin H, Xie J, Xing S, et al. Machine learning-based analysis identifies and validates serum exosomal proteomic signatures for the diagnosis of colorectal cancer. Cell Rep Med. 2024;5(8):101689. IF=10.6 Zone 1 • Document download link: https://www.sciencedirect.com/science/article/pii/S2666379124004105?via%3Dihub Literature download link: https://www.sciencedirect.com/science/article/pii/S2666379124004105?via%3Dihub • 2. Cheng B, Li C, Li J, et al. The activity and immune dynamics of PD-1 inhibition on high-risk pulmonary ground glass opacity lesions: insights from a single-arm, phase II trial. Signal Transduct Target Ther. 2024;9(1):93. IF=52.7 Zone 1 Literature download link: https://www.nature.com/articles/s41392-024-01799-z 2. Cheng B, Li C, Li J, et al. The activity and immune dynamics of PD-1 inhibition on high-risk pulmonary ground glass opacity lesions: insights from a single-arm, phase II trial. Signal Transduct Target Ther. 2024;9(1):93. IF=52.7 Zone 1 Literature download link: https://www.nature.com/articles/s41392-024-01799-z • 3. Min L, Bao H, Bu F, et al. Machine-Learning-Assisted Procoagulant Extracellular Vesicle Barcode Assay toward High-Performance Evaluation of Thrombosis-Induced Death Risk in Cancer Patients. ACS Nano. 2023;17(20):19914-19924. IF=16.0 Zone 1 Literature download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABA0mvAjeb 3. Min L, Bao H, Bu F, et al. Machine-Learning-Assisted Procoagulant Extracellular Vesicle Barcode Assay toward High-Performance Evaluation of Thrombosis-Induced Death Risk in Cancer Patients. ACS Nano. 2023;17(20):19914-19924. IF=16.0 Zone 1 Literature download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABA0mvAjeb • 4. Wang Y, Li X, Wei https://pmc.ncbi.nlm.nih.gov/articles/PMC10507283/ 4. Wang Y, Li X, Wei https://pmc.ncbi.nlm.nih.gov/articles/PMC10507283/ • 5. Guo W, Zhou B, Zhao L, et al. Plasma extracellular vesicle long RNAs predict response to neoadjuvant immunotherapy and survival in patients with non-small cell lung cancer. Pharmacol Res. 2023;196:106921. IF=10.5 Zone 2 Literature download link: https://www.sciencedirect.com/science/article/pii/S1043661823002773?via%3Dihub 5. Guo W, Zhou B, Zhao L, et al. Plasma extracellular vesicle long RNAs predict response to neoadjuvant immunotherapy and survival in patients with non-small cell lung cancer. Pharmacol Res. 2023;196:106921. IF=10.5 Zone 2 Literature download link: https://www.sciencedirect.com/science/article/pii/S1043661823002773?via%3Dihub • 6. Shen S, Wang C, Gu J, et al. A Predictive Model for Initial Platinum-Based Chemotherapy Efficacy in Patients with Postoperative Epithelial Ovarian Cancer Using Tissue-Derived Small Extracellular Vesicles. J Extracell Vesicles. 2024;13(8):e12486. IF=14.5 Zone 1 Literature download link: https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jev2.12486 6. Shen S, Wang C, Gu J, et al. A Predictive Model for Initial Platinum-Based Chemotherapy Efficacy in Patients with Postoperative Epithelial Ovarian Cancer Using Tissue-Derived Small Extracellular Vesicles. J Extracell Vesicles. 2024;13(8):e12486. IF=14.5 Zone 1 Literature download link: https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jev2.12486 • 7. Qin C, Zhao B, Wang Y, et al. Extracellular vesicles miR-31-5p promotes pancreatic cancer chemoresistance via regulating LATS2-Hippo pathway and promoting SPARC secretion from pancreatic stellate cells. J Extracell Vesicles. 2024;13(8):e12488. IF=14.5 Zone 1 Literature download link: https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jev2.12488 7. Qin C, Zhao B, Wang Y, et al. Extracellular vesicles miR-31-5p promotes pancreatic cancer chemoresistance via regulating LATS2-Hippo pathway and promoting SPARC secretion from pancreatic stellate cells. J Extracell Vesicles. 2024;13(8):e12488. IF=14.5 Zone 1 Literature download link: https://isevjournals.onlinelibrary.wiley.com/doi/10.1002/jev2.12488 • 8. Yin D, Wang P, Hao Y, et al. A battery-free nanofluidic intracellular delivery patch for internal organs. Nature. 2025;642(8069):1051-1061. IF=48.5 Zone 1 Download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABAZQmMaVK 8. Yin D, Wang P, Hao Y, et al. A battery-free nanofluidic intracellular delivery patch for internal organs. Nature. 2025;642(8069):1051-1061. IF=48.5 Zone 1 Download link: https://drive.weixin.qq.com/s?k=ADwAeQcoABAZQmMaVK

Documents

ES9P11e说明书.pdf

ES9P14e说明书.pdf

ES9P021e.pdf

ES9P024e.pdf

FAQ

What is Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples?: Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples is an exosome research reagent under the Exosome Purification Kits category. Overview In the minimum experimental requirements (MISEV2023) recommended by the International Society for Extracellular Vesicles (ISEV), the recommendations for isolating EVs based on commercial kits are as follows: Kits based on the principle of precipitati…
What experiments is Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples used for?: Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples is suitable for exosupur ® exosome purification kit is based on molecular size exclusion chromatography SEC for the isolation and purification of small extracellular vesicles sEV. It is suitable for the isolation and purification of exosomes from complex biological samples such as plasma and serum, as well as trace plasma/serum (200µl) complex biological samples. It has the advantages of high purity, robust yield, convenience and speed, and is a robust assistant for exosomes and functional research. This kit provides a systematic solution for the isolation of sEV from complex biological samples ranging from 1 ml of normal plasma/serum to 0.2 ml of trace plasma/serum.
Which product category does Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples belong to?: Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples 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 Exclusion Column kit-Plasma Serum/Micro Complex Samples?: Yes. The page provides ES9P11e说明书.pdf, ES9P14e说明书.pdf, ES9P021e.pdf, ES9P024e.pdf. Detailed specifications should be confirmed with the page or official documents.
What should be confirmed before using Exosupur ® Exosome Purification Exclusion Column kit-Plasma Serum/Micro Complex Samples?: Users should confirm sample type, experimental goal, starting volume or particle concentration, downstream assays, and whether supporting purification, characterization, or tracing workflows are required.