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EasySep™细胞外囊泡PE正选试剂盒

用于从20 mL生物体液或条件培养基中分选经PE标记的细胞外囊泡
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产品号 #（选择产品）

产品号 #100-0812_C

采用免疫磁珠正选技术快速简便地分选细胞外囊泡

产品优势

  • 快速、简便
  • 无需分离柱

产品组分包括

  • EasySep™细胞外囊泡PE正选抗体混合物，1 mL
  • EasySep™ Releasable RapidSpheres™ 50201磁珠，4 x 1 mL
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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

高效获得特定类型的细胞外囊泡（EV），可用于后续分析，或提升生物标志物开发测定的灵敏度。使用EasySep™ 细胞外囊泡PE正选剂盒，仅需 70 分钟即可从多种生物流体或细胞培养条件培养基中分选组织或细胞特异性EV亚型。

EasySep™细胞外囊泡PE正选试剂盒可与您选择的PE标记抗体一起使用。通过免疫磁分选方式，基于EV表面标志物分离特定亚型的EV。与传统的EV分离技术（如差速超速离心法）相比，该方法快速、简便、无柱的方案，能实现更高效的EV亚型富集，从而更容易在下游分析中检测低丰度EV。分选得到的EV保持完整性，可进行下游分析，如 DNA/RNA 提取、Western印迹或质谱分析。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • EasyPlate™ EasySep™ Magnet (Catalog #18102) • “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
 
种属
人
 
筛选方法
Positive
 
应用
细胞外囊泡分选
 
品牌
EasySep
 
研究领域
癌症，细胞外囊泡研究，免疫
 

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Data Figures

Figure 1. Representative Western Blot Analyses of EV Subtypes Isolated from Human and Mouse Plasma

(A) EVs were isolated from healthy human plasma spiked with cancer cell (MCF7) derived EV using PE-conjugated anti-human CD45 (Clone HI30, Catalog # 60018PE), PE-conjugated anti-human EpCAM (Clone 9C4), or PE-conjugated Mouse IgG1 isotype control (Clone MOPC, Catalog # 60070PE) with the EasySep™ Human Extracellular Vesicle PE Positive Selection Kit; or differential ultracentrifugation. Target marker (EpCAM and CD45) and common EV markers (CD9, CD81, and CD63) are analyzed by western blot under non-reducing conditions. (B) EVs were isolated from mouse plasma using PE-conjugated anti-mouse CD63 (Clone NVG-2) or PE-conjugated Rat IgG2a isotype control with the EasySep™ Human Extracellular Vesicle PE Positive Selection Kit, target marker is analyzed by western blot under non-reducing condition.

Figure 2. EasySep™ Immunomagnetic Isolation of EpCAM+ EVs from Complex Biofluids

EpCAM+ EVs were isolated using a biotin-conjugated anti-human EpCAM antibody and the EasySep™ Release Human Biotin Positive Selection Kit. Conditioned media (CM) from breast cancer (CAMA-1 or MCF7) or hepatic organoid culture were serially diluted with phosphate-buffered saline (PBS) to simulate starting EV concentrations typically found in cell culture. Healthy donor plasma was spiked with different concentrations of purified MCF7-derived EVs to simulate cancer patient plasma. EasySep™ immunomagnetic isolation successfully recovered EpCAM+ EVs over a wide range of EV concentrations (10^8 - 10^11 EVs/mL, measured by field flow fractionation coupled with multi-angle light scattering analysis). Western blot densitometry analysis of isolated EVs showed EpCAM protein density increased with the EV concentrations in the samples. This demonstrated EasySep™ isolation coupled with downstream analyses can differentiate samples containing high and low amounts of EpCAM+ EV, such as cancer patients versus healthy donors.

Figure 3. EV Integrity and Cargo are Maintained Following EasySep™ Immunomagnetic Isolation

CD63+ EVs were isolated from plasma using the EasySep™ Human Extracellular Vesicle (CD63) Positive Selection Kit (n = 2); CD45+ EVs were isolated from plasma using a PE-conjugated anti-human CD45 antibody and the EasySep™ Release Human PE Positive Selection Kit (n = 1); EpCAM+ EVs were isolated from MCF7-derived CM using a biotin-conjugated anti-human EpCAM antibody and the EasySep™ Release Human Biotin Positive Selection Kit (n = 1). Positively isolated EVs were either diluted with PBS (“No particle release EV Control” - grey bars) or diluted with release buffer followed by magnetic incubation to remove particles (“Particle-released EV” - orange bars). EV integrity was determined by RNase digestion with or without first lysing EVs using 0.1% Triton™ X-100. EV cargo recovery was analysed by assessing expression levels of Let-7a miRNA using RT-qPCR. EasySep™ technology successfully isolated intact CD63+, CD45+, and EpCAM+ EVs, which protected miRNA cargo from RNase degradation. The particle-released EVs showed similar Let-7a Ct values as the “no particle release” EV control in all conditions tested, confirming the particle release and magnetic removal steps did not negatively impact miRNA recovery and EV integrity.

Figure 4. Improved Specificity and Yield of EVs Using EasySep™ Immunomagnetic Isolation

EVs were isolated from (A) simulated cancer patient plasma (healthy donor plasma spiked with purified MCF7-derived EVs; n = 3) and (B) healthy donor plasma (n = 3). Bulk/Pan EV isolation was done by differential ultracentrifugation (UC) or the EasySep™ Pan-Extracellular Vesicle Positive Selection Kit targeting CD9, CD63, and CD81. Specific EV subtypes were isolated using EasySep™ EV kits that directly target a single marker or indirectly using PE-conjugated antibodies. Isolated EVs were resuspended in the same volume for all subsequent analyses. EV recovery was analyzed by western blot (loaded with equal sample volumes), comparing EV marker density from each sample relative to UC-isolated EVs. EV marker expression relative to UC was calculated by dividing recovery of a single EV marker by the total recovery of all 5 markers. EasySep™ Pan EV isolation recovered more EVs expressing CD9, CD63, and CD81 markers than UC from simulated cancer patient (panel A) and healthy donor samples (panel B). EasySep™ isolations targeting EVs by CD9, CD63, CD81, CD45, or EpCAM recovered distinct EV subtypes with a high expression of the target marker. Specific targeting of CD45 and EpCAM showed the possibility to separate immune cell- and cancer cell-derived EVs from the same patient sample: CD45-targeted EVs expressed CD45 but not EpCAM; EpCAM-targeted EVs expressed EpCAM but not CD45. Although EpCAM+ EVs were recovered from simulated patient plasma using UC and EasySep™ methods, only the EasySep™ EpCAM isolation was able to recover EpCAM+ EVs in 2 out of 3 healthy donors (indicated by *), demonstrating EV subtype enrichment improved detection of low frequency EVs in downstream analyses.

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™ Extracellular Vesicle PE Positive Selection Kit
Catalog #
100-0812
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Extracellular Vesicle PE Positive Selection Kit
Catalog #
100-0812
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Extracellular Vesicle PE Positive Selection Kit
Catalog #
100-0812
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

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更多信息
Species Human
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • EasyPlate™ EasySep™ Magnet (Catalog #18102) • “The Big Easy” EasySep™ Magnet (Catalog #18001)
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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