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EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit

Immunomagnetic positive selection of human EVs from plasma, serum, cell culture conditioned medium, and urine samples

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EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit

Immunomagnetic positive selection of human EVs from plasma, serum, cell culture conditioned medium, and urine samples

From: 479 USD
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Immunomagnetic positive selection of human EVs from plasma, serum, cell culture conditioned medium, and urine samples
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Product Advantages


  • Fast, easy to use, and column-free.

  • Avoid the need for ultracentrifugation and associated time consuming EV isolation methods.

  • Obtain highly purified EVs without contaminating biofluid components.

What's Included

  • EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit (Catalog #17891)
    • EasySep™ Human Pan-Extracellular Vesicle Positive Selection Cocktail 1 x 1 mL
    • EasySep™ Releasable RapidSpheres™ 50201, 2 x 1 mL
    • NOTE: Isolated extracellular vesicles should not be released from the EasySep™ Releasable Rapidspheres™ 50201
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

Overview

Easily isolate highly purified human extracellular vesicles (EVs) from plasma, serum, cell culture conditioned medium, and urine samples, using immunomagnetic positive selection with the EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit. Widely used in published research for more than 20 years, EasySep™ combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.

In this EasySep™ positive selection procedure, desired EVs are labeled with antibody complexes recognizing the specific tetraspanin markers CD9, CD63, and CD81, and magnetic particles. Labeled EVs are separated using an EasySep™ magnet and by simply pouring or pipetting off the unwanted biofluid components. The EVs of interest remain in the tube. Following magnetic EV isolation in less than 30 minutes, the desired EVs are ready for downstream applications such as DNA/RNA extraction, western blot, or mass spectrometry. This method yields EV preparations with low levels of contaminating proteins, such as apolipoproteins and albumin, helping to ensure cleaner samples for sensitive downstream analyses.

Learn more about how immunomagnetic EasySep™ technology works. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
Subtype
Cell Isolation Kits
Cell Type
Other
Sample Source
Other
Selection Method
Positive
Application
Cell Isolation, Extracellular Vesicle Isolation
Area of Interest
Extracellular Vesicle Research

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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