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EasySep™人总细胞外囊泡正选试剂盒

免疫磁珠正选实现人细胞外囊泡快速简便分离
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产品号 #（选择产品）

产品号 #17891_C

免疫磁珠正选实现人细胞外囊泡快速简便分离

产品优势

  • 快速、简单、无柱
  • 避免使用耗时且需要超速离心的方法分离EV
  • 获取高纯度EVs，避免污染生物液体成分

产品组分包括

  • EasySep™人总细胞外囊泡正选试剂盒（产品号 #17891）
    • EasySep™人总细胞外囊泡正选抗体混合物，1 x 1 mL
    • EasySep™ Releasable RapidSpheres™ 50201磁珠，2 x 1 mL
    • 注意：分选的细胞外囊泡不应从EasySep™ Releasable RapidSpheres™ 50201磁珠上解离下来
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总览
实验数据
产品说明书及文档
应用领域
相关材料与文献
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总览

EasySep™人总细胞外囊泡正选试剂盒通过免疫磁珠正选技术，从血浆、血清和条件培养基中分离人细胞外囊泡。细胞外囊泡被识别CD9、CD81和CD63的抗体四聚体复合物与磁珠标记，随后通过EasySep™磁极分离被标记的细胞，无需使用分离柱。不需要的生物液体组分通过简单倾倒弃去，而所需的细胞外囊泡则被保留在试管中。分离的细胞外囊泡可用于下游应用，如DNA/RNA提取、Western blot或质谱分析。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001)
 
亚型
细胞分选试剂盒
 
细胞类型
其它细胞系
 
样本来源
其它细胞系
 
筛选方法
Positive
 
应用
细胞分选，细胞外囊泡分选
 
研究领域
细胞外囊泡研究
 

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Data Figures

Positive isolation of EVs with CD9, CD63, and CD81 tetraspanin markers from human plasma and MSC-conditioned medium.

Figure 1. Typical Western Blot Analyses of EVs Isolated from Human Plasma and Mesenchymal Stromal Cell (MSC)-Conditioned Medium

The Western blot analyses in the above examples show positive isolation of EVs with CD9, CD63, and CD81 tetraspanin markers from (A) human plasma and (B) MSC-conditioned medium. Western blots were run under non-reducing conditions.

Figure 2. Tetraspanin Protein Expression and Recovery of EVs Using EasySep™ Human Pan/CD9/CD63/CD81 Extracellular Vesicle Positive Selection Kits

(A) EVs were isolated from plasma using either EasySep™ Human Extracellular Vesicle Positive Selection Kits or differential ultracentrifugation (2 x 70 min, 100,000 xg) and isolated fractions were analyzed by western blot for tetraspanin protein expression. (B) Equal or higher recovery of EVs was achieved from mesenchymal stromal cell (MSC)-conditioned MesenCult™-ACF Plus Medium and plasma using EasySep™ Human PanExtracellular Vesicle Positive Selection Kit when compared to EVs isolated using other commercially available immunocapture-based EV isolation kits.

Figure 3. Images of Plasma-Derived EVs Isolated Using EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit

Transmission electron microscopy (TEM) analysis following EasySep™ positive selection shows intact and spherical-shaped (arrows) plasma-derived EVs. The isolated EVs are attached to tetrameric antibody complexes and magnetic particles.

Figure 4. Top 10 Cellular Compartment Gene Ontology Terms for EVs Isolated from Plasma or MSC-Conditioned Medium Using EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit

Proteins present in EVs isolated from (A) plasma and (B) MSC-conditioned medium using EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit were detected by proteomic analysis. \Gene Ontology analysis showed the detected proteins were grouped by terms associated with EVs, confirming the quality and compatibility of isolated EVs with mass spectrometry analysis.

Figure 5. Common microRNAs (miRNAs) in Plasma-Derived EVs Isolated Using EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit

MicroRNAs (miRNAs) found in plasma were detected with RT-qPCR demonstrating the compatibility of EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit with downstream RNA extraction and RNA analysis. The increase in Ct value following EV lysis using 0.1% Triton™X-100 and RNase digestion demonstrate the integrity of isolated EVs.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit
Catalog #
17891
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit
Catalog #
17891
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit
Catalog #
17891
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (7)

Isolate and Purify Extracellular Vesicles
Brochure
Isolate and Purify Extracellular Vesicles
Isolating Extracellular Vesicles from Urine with EasySep™ EV Human Positive Selection Kits
Technical Bulletin
Isolating Extracellular Vesicles from Urine with EasySep™ EV Human Positive Selection Kits
How to Characterize Extracellular Vesicles by Western Blotting
Protocol
How to Characterize Extracellular Vesicles by Western Blotting
Mesenchymal Stromal Cell-Derived Exosomes: Biogenesis and Cargoes
Wallchart
Mesenchymal Stromal Cell-Derived Exosomes: Biogenesis and Cargoes
High-Throughput Isolation of Extracellular Vesicles for Next-Generation Diagnostic Assays
25:25
Webinar
High-Throughput Isolation of Extracellular Vesicles for Next-Generation Diagnostic Assays
Decoding Intercellular Communication via Hepatocyte-Secreted Extracellular Vesicles
35:31
Webinar
Decoding Intercellular Communication via Hepatocyte-Secreted Extracellular Vesicles
Online Immunology Journal Club: Viral Exploitation of Extracellular Vesicles to Spread Infection
23:18
Webinar
Online Immunology Journal Club: Viral Exploitation of Extracellular Vesicles to Spread Infection

Publications (1)

A method of separating extracellular vesicles from blood shows potential clinical translation, and reveals extracellular vesicle cargo gremlin-1 as a diagnostic biomarker. N. McNamee et al. Translational oncology 2022 jan

Abstract

Extracellular vesicles (EVs) have potential as minimally invasive biomarkers. However, the methods most commonly used for EV retrieval rely on ultracentrifugation, are time-consuming, and unrealistic to translate to standard-of-care. We sought a method suitable for EV separation from blood that could be used in patient care. Sera from breast cancer patients and age-matched controls (n = 27 patients; n = 36 controls) were analysed to compare 6 proposed EV separation methods. The EVs were then characterised on 8 parameters. The selected method was subsequently applied to independent cohorts of sera (n = 20 patients; n = 20 controls), as proof-of-principle, investigating EVs' gremlin-1 cargo. Three independent runs with each method were very reproducible, within each given method. All isolates contained EVs, although they varied in quantity and purity. Methods that require ultracentrifugation were not superior for low volumes of sera typically available in routine standard-of-care. A CD63/CD81/CD9-coated immunobead-based method was most suitable based on EV markers' detection and minimal albumin and lipoprotein contamination. Applying this method to independent sera cohorts, EVs and their gremlin-1 cargo were at significantly higher amounts for breast cancer patients compared to controls. In conclusion, CD63/CD81/CD9-coated immunobeads may enable clinical utility of blood-based EVs as biomarkers.
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更多信息

更多信息
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001)
Sample Source Other
Selection Method Positive
标记抗体
  1. 细胞外囊泡人CD9/CD63/CD81抗体组合
    细胞外囊泡人CD9/CD63/CD81抗体组合

    使用 CD9、CD63 和 CD81 标记物检测细胞外囊泡的抗体试剂盒

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