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美国NanoView公司所开发的全自动外泌体荧光检测分析系绞/strong>是一款无需纯化的、全自动的可对单个外泌体进行表征分析的全新设备。该设备能够提供全方位的外泌体表征信息,包括外泌体粒径大小、计数、分布、携带蛋白表达、生物标志物(CD9,CD81,CD63等)共定位等。操作简单,结果可靠。一经推出,便引起了外泌体领域科研工作者的广泛关注,短短两年时间在全球已有50多个实验室采用该技术,发表重要文献近百篇、/span>
全自动外泌体荧光检测分析系绞/strong>的基本原理是一种基于特异性免疫捕获技术,允许研究者直接分析特定群体的外泌体或外囊泡。通过配套的试剂盒,客户一次性能够分析多?个不同的样本,大大节省了时间和经济成本、strong>全自动外泌体荧光检测分析系绞/strong>兼容各种生物样本,除了纯化的外泌体之外,对于血液、尿液、恶性肿瘤、腹水中的外泌体也可直接检测分析,大大拓展了研究范围、/span>
应用方向及主要特?/strong>
生物标记物共定位 可量?种标记物的表达情冴/p> | 计数分析 直接从样品中计算抗原阳性外泌体的数量,无需提纯 |
粒径分析 高精度统计外泌体的颗粒大小及分布 | 检测外泌体内容?/strong> 使用ExoView Cargo试剂盒可探测外泌体内部核酸的装载情况,分析装载率 |
荧光检浊/strong> 具备3个荧光通道,能够探测单个蛋白的结合 | 线性工作流稊/strong> 9个样品的全自动分枏/p> |
无须纯化 无需担心纯化带来的误差,更精确的测量样品间的表征和表达信息的差异 | 多重样本分析 可使?种表面标记物来筛选外泌体 |
粒径分析
全自动外泌体荧光检测分析系绞/strong>能够寸50 nm的外泌体进行全方面的表征,无论是粒径尺寸、粒径分布还是外泌体的亚型均可在一次测试中得到。并且所用来测试的样本无需进行纯化,避免因纯化带来的样本偏差、/span>
分析不同大小和不同尺度的外泌体亚群,在CD171过表达系统中?00 nm以上的外泌体仅表达了CD171、/span>
量化外泌体亚羣/strong>
通过抗体捕获的模式,全自动外泌体荧光检测分析系绞/strong>能够量化含有不同标记物的外泌体亚群,并对不同种群的外泌体进行特异性计数和分析。整个过程无需纯化,并且线性范围可跨越3个数量级、/span>
TSPAN8阳性细胞外囊泡的稀释曲线。使?:3比例的梯度稀释。用R2 = 0.9986计算与TSPAN8荧光数据的线性拟合、/span>
检测外囊泡中的装载?/strong>
MISEV建议在表征外泌体和外囊泡时,应当同时测量表面和载体蛋白。使用ExoView芯片可穿透外泌体,探测膜内蛋白和载体。并且单次可定量3种表面、管腔蛋白、/span>
检测细胞外泌体内的Syntenin、/span>
WT细胞在未穿膜的条件下几乎观测不到Syntenin的信号。进行穿膜处理后可以观测到Syntenin信号,而KO之后信号消失
生物标志物共定位
全自动外泌体荧光检测分析系绞/strong>能够在单个外泌体样本上检测多?种标记物,并同时提供外泌体的其它表征数据诸如计数、颗粒尺寸统计等、/span>
无需纯化
全自动外泌体荧光检测分析系绞/strong>只测量含有靶标抗原的特定细胞外泌体群体。仅需35ul的稀释样品,即可直接获得外泌体的表型、l粒径和计数信息。并且无需担心污染物对测试的影响、/span>
无论纯化与否,Exoview均可进行分析
ExoView? Kit 配置清单:
ExoView? Tetraspanin Kits | CD9 - 捕获抖/span>佒/span> CD63 - 捕获抗体 CD81 - 捕获抗体 Mouse IgG 阴性对 CD9、CD63、CD81三色共标记荧光抗体、/span> |
ExoView? Tetraspanin Plasma Kits | CD9 - 捕获抗体 CD63 - 捕获抗体 CD81 - 捕获抗体 CD41a - 捕获抗体 多达三种定制化捕获抗佒/span> 同种阴性对 CD9、CD63、CD81三色共标记荧光抗体、/span> |
ExoView? Tetraspanin Custom Kits | CD9 - 捕获抗体 CD63 - 捕获抗体 CD81 - 捕获抗体 多达三种定制化捕获抗佒/span> 同种阴性对?/span> CD9、CD63、CD81三色共标记荧光抗体、/span> |
ExoView? Cargo Kits | 兼容所有ExoView Kit 标准 CD9 CD63和CD81 捕获抗体 Mouse IgG 阴性对 ExoView Cargo 试剂 CD9、CD63、CD81三色共标记荧光抗体、/span> Sytenin荧光抗体 |
发表文章
2020年已发表文献
1. Cytokine profiling in serum-derived exosomes isolated by different methods Jung HH Kim JY Lim JE Im YH Nature Scientific Reports 2020
2. Study of immune-tolerized cell lines and extracellular vesicles inductive environment promoting continuous expression and secretion of HLA-G from semiallograft immune tolerance during pregnancy Cho K Kook H Kang S Lee J JEV 2020
3. Annexin A1–dependent tethering promotes extracellular vesicle aggregation revealed with single–extracellular vesicle analysis Rogers MA Buffolo F Schlotter F Atkins SK Lee LH Halu A Blaser MC Tsolaki E Higashi H Luther K Daaboul G Bouten CVC Body SC Singh SA Bertazzo S Libby P Aikawa M Aikawa E Cell Biology 2020
4. Targeting tumor-derived exosomes using a lectin affinity hemofiltration device Marleau AM Jacobs MT Gruber N Rodell TC Ferrone S Whiteside TL Cancer Research2020
5. Extracellular Vesicle and Particle Biomarkers DefineMultiple Human Cancers. Hoshino A Kim HS Bojmar L Gyan KE Cioffi M Hernandez J Zambirinis CP Rodriques G Molina H Heissel S Mark MT Steiner L Benito Martin A Lucotti S Di Giannatale A Offer K Nakajima M Williams C Lyden D. Cell 2020
6. Reporter mice for isolating and auditing cell type‐specific extracellular vesicles in vivo McCann JV Bischoff SR Zhang Y Cowley DO Sanchez-Gonzalez V Daaboul GG Dudley AC Genesis2020
7. High yield and scalable EV production from suspension cells triggered by turbulence in a bioreactor Grainger A Wilhelm C Gazeuah F Silva A Cytotherapy 2020
8. Extracellular Vesicles: A New Frontier for Research in Acute Respiratory Distress Syndrome Mahida RY Matsumoto S Matthay MA. American Journal of Respiratory Cell and Molecular Biology2020
9. Small extracellular vesicles modulated by V3 integrin induce neuroendocrine differentiation in recipient cancer cells Quaglia F Krishn SR Daaboul GG Sarker S Pippa R Domingo-Domenech J Kumar G Fortina P McCue P Kelly WK Beltran H Liu Q Languino LR. Journal of Extracellular Vesicles Volume 9 Issue 1 2020
10. Characterisation of extracellular vesicles surface markers and co-expression studies with single particle interferometric imaging platform Kusuma G Lim R. Cytotherapy Volume 22 Issue 5 2020
11. Engineering mesenchymal stem cell paracrine activity with 3D culture Kusuma G LiA Zhu D McDonald H Chmabers D Frith J Lim R.+strong>Cytotherapy Volume 22 Issue 5 2020
12. Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria,Ahn JY Datta S Cano M Mallick E Rai U Powell B Tian J Witwer KW Handa JT Paulaitis ME.+strong>Biochim Biophys Acta Gen Subj. 2020 Mar 30:129598. (doi: 10.1016/j.bbagen.2020.129598)
13. Advantageous Antibody Microarray Fabrication Through DNA-Directed Immobilization: A Step Toward Use of Extracellular Vesicles in Diagnostics Brambila D Sola L Chiari M. Talanta 2020
15. Membrane-Binding Peptides for Extracellular Vesicles On-Chip Analysis Gori A Romanato A Bergamaschi G Strada A Gagni P Frigerio R Brambilla D Vago R Galbiati S PIcciolini S Bedoni M Daaboul G Chiari M Cretich M. Journal of Extracellular Vesicles Volume 9 Issue 1 2020
16. Subpopulations of extracellular vesicles from human metastatic melanoma tissue identified by quantitative proteomics after optimized isolation Crescitelli R L?sser C Jang SC Cvjetkovic A Malmh?ll C Karimi N H??g JL Johansson I Fuchs J Thorsell A Gho YS Bagge RO L?tvall J Journal of Extracellular Vesicles 2020
17. Extracellular Vesicles Derived from Induced Pluripotent Stem Cells Promote Renoprotection in Acute Kidney Injury Model Collino F Lopes JA Tapparo M Tortelote GG Kasai-Brunswick TH Lopes GMC Almeida DB Skovronova R Wendt CHC de Miranda K Bussolati B Vieyra A Soares Lindoso R. Cells 2020
18. Extracellular Vesicles: A New Frontier for Research in Acute Respiratory Distress Syndrome,Mahida RY Matsumoto S Matthay MA. American Journal of Respiratory Cell and Molecular Biology 2020
19. Exosomes A clinical COmpendium - Methods for exosome isolation and characterization,Zhou M Weber SR Zhao Y Chan H Sundstrom JM. Exosomes A Clinical Compendium 2020
20. Phenotypic analysis of extracellular vesicles: a review on the applications of fluorescence,Panagopoulou MS Wark AW Birch DJS Gregory CD. Journal of Extracellular Vesicles 2020
21. Immune Cell-Derived Exosomes in the Cancer-Immunity Cycle Yan W Jiang S Trends in Cancer 2020
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如何使用全自动外泌体荧光检测分析系绞/strong>ExoView进行外泌体测量?
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