Exosome是由多种活细胞分泌的50-100nm的膜性小囊泡,来源于晚期核内体(也称为多囊泡体)的囊泡,是由细胞内多泡体(MVBs)出芽形成,再与细胞膜融合后释放到细胞外基质中。几乎所有类型的细胞都能分泌,广泛存在于各种体液中。不同细胞来源的Exosome所含有的具体成分不尽相同,根据分泌的细胞类型不同,在血液、尿液、羊水、恶性肿瘤、腹水等动物体液中的Exosome通常含有特定的miRNA和蛋白质。因此,Exosome为研究潜在的各种biomarker提供了很好的生物学材料。Exosome参与调控重要的细胞生理活动,在**应答、凋亡、血管生成、炎症反应、凝结过程中的作用均有报道,可以成为多种**的早期诊断标记物,也能作为靶向**的载体进行****。作为囊泡的一种,Exosome受到越来越多的关注。其分泌过程如图[1]:
SBI公司Exosome外泌体一体化研究工具一览
SBI公司是一家提供*先进的供**外泌体研究所需产品和试剂盒,包括分离,检测和定量试剂盒,探测(表征和分析试剂盒),为广大研究者提供了*新的研究手段。2009年,SBI公司的研究团队们发现了Exosome的巨大研究价值后开发了**款商业化的外泌体分离试剂盒,期间也一直在壮大Exosome研究团队,提升产品质量丰富产品种类成为世界上*大的Exosome研究供应商。SBI将会把*新的技术和方法转化成加快您外泌体研究进程的*有力的工具。
外泌体的提取
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分离完整的Exosome
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FACS磁珠分选
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IP高通量**沉淀Exosome
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不含Exosome的培养基
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Exosome的验证分析
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抗体
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Elisa定量分析
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CETP活性定量分析
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Exosome RNA的研究
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RNA的提取
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RNA二代测序建库试剂盒
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Exosome功能研究
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荧光标记
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转染试剂
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Ev-Entry system
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X PACK 技术
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XMIR miRNA packing system
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XStamp Exosome Targeting Technology
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一 外泌体的分离
SBI公司研究和开发了一款基于多聚体形式的Exosome提取试剂,能够快速有效地提取及分离血清和肿瘤组织样本中的Exosome囊泡。这些多聚体能够形成类似于网状结构,从而将60-180nm直径的微泡分离出来。其纯度和产量皆优于传统方法。
(一)外泌体分离纯化产品选择
分离完整Exosome
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ExoQuick:
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适合富含Exosome样本如:血清、腹水
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ExoQuick-TC:
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适合Exosome含量较少的样本如:细胞培养上清、尿液、脑脊液
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ExoQuick Plasma Kit:
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血浆(试剂盒中含Thrombin及ExoQuick试剂)
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ExoQuick-CG:
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药品生产管理规范标准级别的用于临床前的应用研究(适用于各种培养液、体液如尿液、唾液、脑脊液等等样本的Exosome提取)
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ExoQuick-LP
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预处理血清或血浆样本,以出去脂蛋白
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产品信息SBI公司Exosome外泌体一体化研究工具一览
货号
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产品
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规格
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价格(元)
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EXOQ5A-1
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ExoQuickExosome precipitation solution
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5ml(75T)
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4203.2
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EXOQ20A-1
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ExoQuickExosome precipitation solution
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20ml(300T)
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13561
|
EXOTC10A-1
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ExoQuick TC Exosome precipitation solution
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10ml(10T)
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4189
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EXOTC50A-1
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ExoQuick TC Exosome precipitation solution
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50ml(50T)
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14072
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EXOCG50A-1
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ExoQuick-CG™ Exosome Precipitation Solution
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50ml(30R)
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询价
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EXOQ5TM-1
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ExoQuick Plasma prep and Exosome Precipitation Kit
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75T
|
9471.4
|
关联产品
货号
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产品
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规格
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价格(元)
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TMEXO-1
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Thrombin Plasma prep for Exosome Precipitation
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100T
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6347.4
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EXOLP5A-1
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ExoQuick-LP Lipoprotein PreClear&Exosome Precipitation Kit
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10T
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5751
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(二)FACS磁珠分选Exosomes
SBI为您推荐Exo-Flow抗体磁珠试剂盒,该试剂盒能够捕获Exosome特定的表面标记蛋白,利用流式分选技术,从而分离出不同亚型的Exosome。试剂盒中提供的9.1μM直径的Exo-Flow磁珠与传统的4μM Dynabeads磁珠相比,能够更好地捕获Exosome,特别适用于数量较少的Exosome样本,并且有多种标记抗体供选择。
产品信息
货号
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产品
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规格
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价格(元)
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EXOFLOW100A-1
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CD9 Exo-Flow capture kit (Magnetic streptavidin beads, CD9-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
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5736.8
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EXOFLOW150A-1
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TetraspaninExo-Flow combo capture kit (Magnetic streptavidin beads, CD9, CD63, and CD81-biotin capture antibodies, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
|
8718.8
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EXOFLOW200A-1
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CD31 Exo-Flow capture kit (Magnetic streptavidin beads, CD31-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
|
10 reactions
|
5736.8
|
EXOFLOW210A-1
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CD44 Exo-Flow capture kit (Magnetic streptavidin beads, CD44-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
|
10 reactions
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5736.8
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XOFLOW300A-1
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CD63 Exo-Flow capture kit (Magnetic streptavidin beads, CD63-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
|
5736.8
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EXOFLOW400A-1
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CD81 Exo-Flow capture kit (Magnetic streptavidin beads, CD81-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
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5736.8
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EXOFLOW500A-1
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Rab5b Exo-Flow capture kit (Magnetic streptavidin beads, Rab5b-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
|
5736.8
|
EXOFLOW600A-1
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HLA-G Exo-Flow capture kit (Magnetic streptavidin beads, HLA-G-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
|
10 reactions
|
5736.8
|
EXOFLOW610A-1
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CD14 Exo-Flow capture kit (Magnetic streptavidin beads, CD14-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
|
5736.8
|
EXOFLOW620A-1
|
CD68 Exo-Flow capture kit (Magnetic streptavidin beads, CD68-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
|
5736.8
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EXOFLOW630A-1
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EpCAMExo-Flow capture kit (Magnetic streptavidin beads, EpCAM-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
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10 reactions
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5736.8
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EXOFLOW650A-1
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Alpha-2-Macroglobulin (A2M) Exo-Flow capture kit (Magnetic streptavidin beads, A2M-biotin capture antibody, Wash and Elution Buffers, Exo-FITC stain)
|
10 reactions
|
5736.8
|
CSFLOWBASICA-1
|
Basic Exo-Flow kit without any coupled antibodies (Uncoupled magnetic streptavidin beads, Wash and Elution Buffers, Exo-FITC stain)
|
100 reactions
|
9869
|
关联产品
货号
|
名称
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规格
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价格
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EXOFLOW700A-1
|
Exo-Flow Multifunctional Magnetic Stand
|
1 Kit
|
3947.6
|
EXOFLOW800A-1
|
Exo-FITC Exosome FACS stain
|
20 reactions
|
2158.4
|
EXOFLOW810A-1
|
Exo-APC Exosome FACS stain
|
20 reactions
|
2158.4
|
(三)IP高通量**沉淀Exosomes SBI公司Exosome外泌体一体化研究工具一览
Exo-Flow 96/32 IP试剂盒可从血浆样本、使用ExoQuick-TC或超速离心富集的细胞培养上清(尿液、脑脊髓液等)Exosomes产物样本中高通量、直接**沉淀Exosomes。试剂盒中的磁珠已经预结合了CD9、CD63或者CD81用于选择性捕获特定的Exosomes亚型,直接将上述样本添加入磁珠中即可**沉淀Exosomes。96孔板或32孔板形式产品可用于同时处理多个样本。得到Exosomes具有完整形态,可用于下游Exosomes RNA和蛋白分析。
产品信息
货号
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名称
|
规格
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价格
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EXOFLOW32A-CD9
|
Exo-Flow 32 CD9 IP kit
|
32 rxns
|
询价
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EXOFLOW32A-CD63
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Exo-Flow 32 CD63 IP kit
|
32 rxns
|
询价
|
EXOFLOW32A-CD81
|
Exo-Flow 32 CD81 IP kit
|
32 rxns
|
询价
|
EXOFLOW96A-CD9
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Exo-Flow 96 CD9 IP kit
|
96 rxns
|
询价
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EXOFLOW96A-CD63
|
Exo-Flow 96 CD63 IP kit
|
96 rxns
|
询价
|
EXOFLOW96A-CD81
|
Exo-Flow 96 CD81 IP kit
|
96 rxns
|
询价
|
关联产品
货号
|
名称
|
规格
|
价格
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EXOFLOWMAG-1
|
ExoFlowMag96 96-well plate magnet
|
1 Kit
|
询价
|
二 Exosome验证分析
(一)ELISA定量分析
SBI公司研发的ExoELISA试剂盒适用于所有类型细胞分泌的任何方法分离得到的Exosome,并且有三种Exosome表面特殊蛋白CD63,CD9,CD81供选择。
产品信息
货号
|
产品名称
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规格
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价格(元)
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EXOEL-CD63A-1
|
Exosome ELISA Complete Kit (CD63 detection)
|
96 reactions
|
8378
|
EXOEL-CD81A-1
|
Exosome ELISA Complete Kit (CD81 detection)
|
96 reactions
|
8378
|
EXOEL-CD9A-1
|
Exosome ELISA Complete Kit (CD9 detection)
|
96 reactions
|
8378
|
关联产品
货号
|
产品名称
|
规格
|
价格(元)
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EXOEL96P-1
|
96 Well ExoELISA Plate (12×8well strips,pack of plates)
|
10 Plates
|
5609
|
EXOEL-SD-1
|
Exosome calibration Elisa standards for CD63,CD9 and CD81 ExoELISAs (15 standard curves)
|
1ml
|
询价
|
EXOEL-BB-20
|
Exosome binding buffer for ELISAs
|
20 ml
|
1902.8
|
(二)CETP活性定量分析
SBI公司已通过研究证明,对于血清、干细胞、肿瘤细胞系、Msc来源的Exosome中富含AchE(乙酰辅酶A乙酰胆碱酯)通过检测AchE的活性可直接对Exosome进行定量。
产品信息
货号
|
名称
|
规格
|
价格(元)
|
EXOCET96A-1
|
EXOCET Exosome Quantitation Kit
|
96 rxns
|
询价
|
三 Exosome RNA**研究
(一)RNA的提取
Exosome RNA提取试剂盒专为Exosome RNA抽提设计,**的富集microRNA和其他小片段RNA。分别针对血液样本和体液样本设计两款RNA提取试剂盒,特异性更高,适用所有类型细胞分泌的,以及任何分离方法得到的Exosome RNA提取。操作简单、方便,1h完成血清Exosome RNA提取,无需苯酚/氯仿抽提。纯化的Exosome RNA可以做进一步的microRNA筛选、芯片或二代测序等下游分析。但是由于Exosome RNA含量总体水平偏低,因此需在进行下游分析之前进行Exosome RNA的预扩增。
产品信息
货号
|
产品名称
|
规格
|
价格(元)
|
RA808A-1
|
SeraMirExosome RNA Purification Column kit (20 exoRNA columns)
|
20 preps
|
4189
|
RA806A-1
|
SeraMirExosome RNA Purification only kit for Media and Urine (5ml ExoQuick and 20 exoRNA columns)
|
20 preps
|
询价
|
RA806TC-1
|
SeraMirExosome RNA Purification only kit for Media and Urine (10ml ExoQuick-TC and 10 exoRNA columns)
|
10 preps
|
6006.6
|
关联产品
货号
|
产品名称
|
规格
|
价格(元)
|
RA800A-1
|
Complete SeraMirExosome RNA Amplification kit (5ml ExoQuick, 20 exoRNA columns, cDNA synthesis and amplification components)
|
20 reactions
|
10280.8
|
RA800TC-1
|
Complete SeraMirExosome RNA Amplification kit from Media and Urine (10ml ExoQuick-TC, 10 exoRNA columns, cDNA synthesis and amplification components)
|
10 preps
|
7227.8
|
RA810A-1
|
Human SeraMirExosome RNA 384 microRNA qPCR Profiler Assay Set
|
20 profiles
|
14824.8
|
RA820A-1
|
Human Complete SeraMirExosome RNA Amplification and Profiling Kit (contains cat# RA800A-1, RA805A-1, RA810A-1 components)
|
20 profiles
|
14824.8
|
RA820TC-1
|
Human Complete SeraMirExosome RNA Amplification and Profiling Kit for Media and Urine (contains cat# RA800TC-1 with 50ml ExoQuick-TC, RA805A-1 and RA810A-1 components)
|
20 profiles
|
18644.6
|
RA811A-1
|
Mouse SeraMirExosome RNA 384 microRNA qPCR Profiler Assay Set
|
20 profiles
|
5708.4
|
RA821A-1
|
Mouse Complete SeraMirExosome RNA Amplification and Profiling Kit (contains cat# RA800A-1, RA805A-1, RA811A-1 components)
|
20 profiles
|
14824.8
|
RA821TC-1
|
Mouse Complete SeraMirExosome RNA Amplification and Profiling Kit for Media and Urine (contains cat# RA800TC-1 with 50ml ExoQuick-TC, RA805A-1 and RA811A-1 components)
|
20 profiles
|
18644.6
|
RA812A-1
|
Rat SeraMirExosome RNA 384 microRNA qPCR Profiler Assay Set
|
20 profiles
|
5708.4
|
RA822A-1
|
Rat Complete SeraMirExosome RNA Amplification and Profiling Kit (contains cat# RA800A-1, RA805A-1, RA812A-1 components)
|
20 profiles
|
14824.8
|
RA822TC-1
|
Rat Complete SeraMirExosome RNA Amplification and Profiling Kit for Media and Urine (contains cat# RA800A-1, RA805A-1, RA812A-1 and EXOTC50A-1 components) 20 profiles
|
20 profiles
|
18644.6
|
RA805A-1
|
SeraMir Control spike-in Small RNA and qPCR assay
|
100 assays
|
2215.2
|
RA800P-3
|
SeraMir 3-Universal Primer bonus pack (for an additional 10,000 qPCR reactions)
|
1.2 ml
|
1533.6
|
(二)RNA的二代测序建库
SBI提供高度灵敏的Exosome RNA解决方案,适用于IIlumina测序平台,试剂盒针对Exosome样本特别设计,尤其适合低浓度exoRNA的高灵敏度建库,*适于起始量在1-10ng的exoRNA建库。
产品信息:
货号
|
产品名称
|
规格
|
价格(元)
|
XRNA101A-1
|
XRNA Exosome RNA-Seq NGS library prep kit (index set 1-8)
|
8rxns
|
询价
|
XRNA101A-1
|
XRNA Exosome RNA-Seq NGS library prep kit (index set 9-16)
|
8rxns
|
询价
|
四、Exosome功能研究
(一)荧光标记Exosome
A Exo-Glow Flurescent Label
SBI新产品Exo-Glow试剂盒,能够荧光标记内源的Exosome RNAs为红色,标记内部Exosome蛋白为绿色,从而达到使用荧光显微镜实时追踪Exosome细胞内相互作用和物质运输。
产品信息:
货号
|
产品名称
|
规格
|
价格(元)
|
EXOR100A-1
|
Exo-Red Exosome RNA fluorescent label
|
20 rxns
|
4047
|
EXOG200A-1
|
Exo-Green Exosome protein fluorescent label
|
20 rxns
|
4047
|
EXOC300A-1
|
Exo-Red + Exo-Green Exosome cargo fluorescent label Combo Pack (10 rxns of each label)
|
20 rxns
|
4615
|
B 利用Cyto-Trace让细胞分泌带有荧光的Exosome
跨膜蛋白CD9、CD63和CD81蛋白为Exosomes常见的生物标记蛋白。通过CD63和CD9 Cyto-Trace质粒,可以让您研究的靶细胞分泌带有绿色或者红色荧光的Exosomes,以便下游功能性运输研究。该产品可实时追踪蛋白的运输,用于蛋白的共定位研究。
产品信息
货号
|
产品名称
|
规格
|
价格(元)
|
CYTO120-PA-1
|
pCT-CD63-GFP (pCMV, Exosome/Secretory, CD63 Tetraspanin Tag)
|
10ug
|
7540.2
|
CYTO120R-PA-1
|
pCT-CD63-RFP (pCMV, Exosome/Secretory, CD63 Tetraspanin Tag)
|
10 ug
|
7540.2
|
CYTO122-PA-1
|
pCT-CD9-GFP (pCMV, Exosome/Secretory, CD9 Tetraspanin Tag)
|
10 ug
|
7540.2
|
CYTO123-PA-1
|
pCT-CD9-RFP (pCMV, Exosome/Secretory, CD9 Tetraspanin Tag)
|
10 ug
|
7540.2
|
CYTO124-PA-1
|
pCT-CD81-GFP (pCMV, Exosome/Secretory, CD81 Tetraspanin Tag)
|
10 ug
|
7540.2
|
CYTO125-PA-1
|
pCT-CD81-RFP (pCMV, Exosome/Secretory, CD81 Tetraspanin Tag)
|
10 ug
|
7540.2
|
(二)Exosome转染试剂
Exo-Fect是一种可以将核酸直接转移至分离的到的外泌体中的核酸转染试剂,可将si/miRNA,mRNA,甚至质粒DNA转染Exosome后进入靶细胞。操作过程中,只需将核酸与Exo-Fect试剂和分离得到的Exosome混合,形成运载体。整个过程不超过1小时即可将核酸高效转至Exosome中。
产品信息
货号
|
产品名称
|
规格
|
价格(元)
|
EXFT10A-1
|
Exo-FectExosome Transfection Kit
|
10rxns
|
2769
|
EXFT20A-1
|
Exo-FectExosome Transfection Kit
|
20rxns
|
4970
|
(三)EV-Entry System
胞外囊泡(EVs)和Exosome是细胞间通讯的重要方式,用于运输RNA和蛋白质的纳米级载体。EVs可为靶细胞运输外源性物质,EV-Entry system不仅能够增强细胞摄取EVs的效率,同时还可提高提高EVs释放物质至靶细胞细胞质的效率。EV-Entry system由A,B两种试剂组成,使用时只需将A,B两种试剂和Exosome或者EVs混合即可。整个过程的操作不超过一小时即可完成使EVs或Exosome高效地将物质运输至靶细胞。
产品信息
货号
|
产品名称
|
规格
|
价格
|
EVEN105A-1
|
EV-Entry Reagent System Kit
|
5 rxns
|
询价
|
EVEN110A-1
|
10 rxns
|
询价
|
(四)XPack Exosome Protein Engineering
XPack技术可将任一蛋白包装至Exosome中,并高效地运输至靶细胞中发挥其生物学作用。SBI公司优化了一特殊的多肽序列可将蛋白靶向介导至Exosome膜上,并将该蛋白包装至Exosome分泌至细胞外。XPack多肽序列已经整合在了XPack克隆和慢病毒表达载体中,可以使报告蛋白或任一兴趣蛋白包装至Exosome。这一“XPack”多肽序列已经包含在XPack慢病毒表达载体系列中选择合适的XPack reporter Ientivectors 或者pre-loaded reporter Exosomes,可实现通过荧光或者发光反应来动态观察Exosomes。此外,研究者可改造含有兴趣的Exosome,并研究Exosome的靶细胞运输。
产品信息
产品编号
|
产品名称
|
规格
|
价格(元)
|
XPAK510PA-1
|
XPack CMV-XP-MCS-EF1-Puro Cloning Lentivector
|
10ug
|
询价
|
XPAK530PA-1
|
XPack CMV-XP-GFP-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
XPAK531PA-1
|
XPack CMV-XP-RFP-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
XPAK532PA-1
|
XPack CMV-XP-Luciferase-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
XPAK710PA-1
|
XPack MSCV-XP-MCS-EF1-Puro Cloning Lentivector
|
10ug
|
询价
|
XPAK730PA-1
|
XPack MSCV-XP-GFP-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
XPAK731PA-1
|
XPack MSCV-XP-RFP-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
XPAK732PA-1
|
XPack MSCV-XP-Luciferase-EF1-Puro Expression Lentivector
|
10ug
|
询价
|
(五)制备包含目的miRNA的Exosome
不同细胞类型分泌的Exosome含有不同的RNA和蛋白,对于其生物功能的研究很有意义。将特异性的RNA序列选择性包装至Exosome是研究的热点。SBI已经研究了一特殊RNA序列标签能将小RNA包装至Exosome中并分泌至细胞外。XMotif RNA序列标签已经整合到了miRNA和anti-miRNA寡核苷酸中,并且已经构建至载体,将介导miRNA包装至Exosome,这一技术为研究Exosome介导的潜在**作用的miRNAs或anti-miRNAs运输至**细胞的功效,提供了有力工具。
(六)靶向运输至特定细胞
SBI开发了“XStamp”技术即Exosome表面呈现系统可将兴趣蛋白高效地定位于改造过的Exosome表面。该技术基于MFG-E8的C1C2结构域的C末端融合。与XStamp标签融合的蛋白质序列会在分泌的Exosome表面有效展现能够与之融合的蛋白配体。该技术能够将Exosome表面带上靶细胞的位置标签,并引导Exosome将物质运输至正确靶细胞。
产品信息 SBI公司Exosome外泌体一体化研究工具一览
产品编号
|
产品名称
|
规格
|
价格(元)
|
XSTP710PA-1
|
XStamp cloning and expression lentivector MSCV-Leader-MCS-C1C2-EF1-Puro
|
10 ug
|
询价
|
XSTP720PA-1
|
XStamp MLN Exosome Motilin receptor GI tract targeting lentivector
|
10 ug
|
询价
|
XSTP721PA-1
|
XStamp NCAM Exosome brain targeting lentivector
|
10 ug
|
询价
|
XSTP722PA-1
|
XStamp BHP1 Exosome brain homing peptide targeting lentivector
|
10 ug
|
询价
|
XSTP723PA-1
|
XStamp GE11 Exosome EGFR targeting lentivector
|
10 ug
|
询价
|
XSTP724PA-1
|
XStamp Her2 ligand Exosome Her2 receptor targeting lentivector
|
10 ug
|
询价
|
XSTP725PA-1
|
XStamp CD40L Exosome CD40 receptor targeting lentivector
|
10 ug
|
询价
|
XSTP726PA-1
|
XStamp IL2 Exosome IL-2 receptor targeting lentivector
|
10 ug
|
询价
|
文献引用:
[1] Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends.[J]. Journal of Cell Biology, 2013, 200(4):373-383.