Make more informed purchasing decisions with our new product availability and delivery estimate feature, now available on all product pages, in your cart, and during checkout.
Sign In
New to STEMCELL?
Register for an account to quickly and easily purchase products online and for one-click access to all educational content.
Thank you for your interest in this product.
Please provide us with your contact information and your local representative
will contact you with a customized quote. Where appropriate, they can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
By submitting this form, you are providing your consent to STEMCELL Technologies Canada Inc. and its subsidiaries and affiliates (“STEMCELL”) to collect and use your information, and send you newsletters and emails in accordance with our privacy policy. Please contact us with any questions that you may have. You can unsubscribe or change your email preferences at any time.
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.
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.
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.
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.
Antibody kit for the detection of extracellular vesicles using CD9, CD63, and CD81 markers
Item added to your cart
EasySep™ Human Pan-Extracellular Vesicle Positive Selection Kit
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.