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EasySep™细胞外囊泡PE正选试剂盒

对来源于血浆、血清、尿液及细胞培养上清的 PE 偶联抗体标记人细胞外囊泡（EVs）进行免疫磁珠正选分离

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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
相关产品

总览

使用EasySep™细胞外囊泡PE正选试剂盒，通过免疫磁性正选法，可从血浆、血清、尿液和细胞培养上清中分离经藻红蛋白（PE）偶联抗体标记的高纯度人细胞外囊泡（EVs）。EasySep™技术在已发表研究中被广泛应用超过20年，将单克隆抗体的特异性与无柱磁系统的简便性相结合。

在此正选步骤中，EVs通过识别特异性标志的PE偶联抗体（不包含在是集合中）和磁性颗粒进行标记。使用EasySep™磁体分离后，生物液体成分被倾倒或移除，目标EVs保留在试管中。整个过程短至70分钟，所得EVs可直接用于DNA/RNA提取、免疫蛋白印迹或质谱分析。

了解更多关于免疫磁性EasySep™技术的工作原理。探索更多产品，包括培养基、添加剂、抗体等，为您的实验流程提供优化方案。

磁极兼容性
• EasySep™磁极（产品号 #18000）
• EasyPlate™ EasySep™磁极（产品号 #18102）
• “The Big Easy” EasySep™磁极（产品号 #18001）
 
分类
细胞分选试剂盒
 
细胞类型
其他物种
 
种属
人
 
分选方法
正选
 
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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实验数据

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.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

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

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area
Workflow Stages

更多信息

更多信息
物种 人类
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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