Exosome-Human CD9 Isolation Reagent (from cell culture) - FAQs

可以。请参见此海报（https://tools.thermofisher.com/content/sfs/posters/Exosome-poster-ISEV-2013-Boston.pdf）。

此外，这里还有一些相关引用：
•Blood 91:2573 (1998)
•Science 289:444 (2000)
•J Physiol 537:537 (2001)
•Mol Cell Proteomics 12:587 (2013)
•Biol Reprod 81:717 (2009)

是的，基于不同的膜表面标志物所分离外泌小体中的蛋白表达谱之间会有明显差异。这一结论由Tauro等（http://www.ncbi.nlm.nih.gov/pubmed/23230278）的研究证实，该研究团队基于EpCAM或A33这两种膜表面标志物，从人癌细胞系的条件培养基中分离出两群明显不同的外泌小体。蛋白组学研究结果显示这两群外泌小体是独特的。

我们拥有多款外泌小体分离试剂盒，包括外泌小体—人源CD63（货号10606D），外泌小体—人源CD9（货号10614D），外泌小体—人源CD81（货号10616D）和外泌小体—人源EpCAM，适用于凭借这些常用的外泌小体膜表面抗原来实现外泌小体分离操作。如果您希望使用您的自备抗体通过其他特异性膜表面标志物来分离外泌小体，您也可使用我们的Dynabeads外泌小体免疫沉淀（ProteinA，货号10610D），Dynabeads外泌小体免疫沉淀（ProteinG，货号10612D）或用于分离/检测的外泌小体—链霉亲和素产品（货号10608D）。此外，用户也可选用抗小鼠IgG的Dynabeads磁珠（货号11031或11033）和识别特定膜表面标志物的小鼠单抗来实现外泌小体分离操作。

一般通过流式细胞仪（使用CD9、CD63、TSG101和Alix等膜表面标志物）来鉴定外泌小体，通过EM来研究其形态和尺寸，或凭借LC-MS/MS来实现更为详细的蛋白分析。

这些标志物要基于外泌小体的细胞来源进行选择。最常用于外泌小体分离和鉴定的膜表面标志物为CD9、CD63、CD81或TSG101。下表列举了一些最近用于鉴定或分离外泌小体的参考文献和膜表面标志物：

膜表面标志物

参考文献
Alix, CD63, EpiCam, HSP70, TSG101
Mol Cell Proteomics 12:587 (2013)

CD9, CD63
Hum Mol Genet 21:R125 (2012)

CD63, MHC II
J Biol Chem 278:52347 (2003)

CD9, CD81, Lamp1, TSG101
Cancer Res 67:7458 (2007)

CD63
Nature Cell Biol 9:654 (2007)

Alix, CD37, CD53, CD63, CD81, CD82, TSG101
J Cell Biol 200:373 (2013)

CD59, CD63, CD133, TSG101
FASEB J 23:1858 (2009)

除沉淀法之外，外泌小体还可通过超速离心或密度梯度分离法实现分离。用户也可使用靶向外泌小体标志物（例如人源的CD9、CD63、CD81、EpCAM）的Dynabeads磁珠或二抗包被的Dynabeads磁珠（使用靶向其他外泌小体膜表面标志物的不同抗体），凭借磁性方法来分离外泌小体。

外泌小体被报道具有多种不同的功能，如抗原呈递、凋亡、血管发生、炎症和凝血作用，这些作用是通过蛋白/脂类交换或信号途径的激活来实现的。外泌小体提供了一种胞间遗传物质交换的全新机制，并能够介导细胞间的相互通讯。外泌小体也能够转运和传播传染性物质，如朊蛋白和逆转录病毒。

外泌小体是微小的卵形或杯形膜结构，大小在30-150 nm，其中包含有mRNA，microRNA，蛋白和脂类。外泌小体可由正常，异常或肿瘤细胞释放进入血、尿、唾液和乳汁等体液中。外泌小体源于内吞型细胞器，并作为与质膜融合的多泡体（MVB）而从细胞释放出来(J Cell Biol 200:373 (2013)).

Yes. See this poster (https://tools.thermofisher.com/content/sfs/posters/Exosome-poster-ISEV-2013-Boston.pdf).

In addition, here are some citations:
- Blood 91:2573 (1998)
- Science 289:444 (2000)
- J Physiol 537:537 (2001)
- Mol Cell Proteomics 12:587 (2013)
- Biol Reprod 81:717 (2009)

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Yes, exosomes isolated with different surface markers can be distinctive in their protein profile. This has been demonstrated by Tauro et al. (http://www.ncbi.nlm.nih.gov/pubmed/23230278), who isolated two distinctive populations of exosomes based on surface markers EpCam or A33 from conditioned cell culture medium from a human carcinoma cell line. This proteomics study indicated that these two populations of exosomes are unique.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

We have exosome isolation kits for Exosome-Human CD63 (Cat. No. 10606D), Exosome-Human CD9 (Cat. No. 10614D), Exosome-Human CD81 (Cat. No. 10616D), and Exosome-Human EpCAM intended for isolating exosomes with these commonly used exosome surface antigens. If you are interested to isolating exosomes with other specific surface markers using your own antibody, you can use our Dynabeads exosome immunoprecipitation (Protein A, Cat. No. 10610D), Dynabeads exosome immunoprecipitation (Protein G, Cat. No. 10612D), or Exosome-Streptavidin for isolation/detection (Cat. No. 10608D). In addition, anti-mouse IgG Dynabeads magnetic beads (Cat Nos. 11031 or 11033) also can be employed in exosome isolation using mouse monoclonal antibodies against selected surface markers.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Exosomes are usually characterized by flow cytometry (using surface markers such as CD9, CD63, TSG101, and Alix), by EM to study morphology and size, or by detailed protein analysis by LC-MS/MS.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

It depends on the cell source from which the exosomes were derived. The most commonly used surface markers for isolating and characterizing exosomes are CD9, CD63, CD81, or TSG101. Here are some of the recent references and surface markers for identifying or isolating exosomes:

Alix, CD63, EpiCam, HSP70, TSG101 Mol Cell Proteomics 12:587 (2013)
CD9, CD63 Hum Mol Genet 21:R125 (2012)
CD63, MHC IIJ Biol Chem 278:52347 (2003)
CD9, CD81, Lamp1, TSG101 Cancer Res 67:7458 (2007)
CD63 Nature Cell Biol 9:654 (2007)
Alix, CD37, CD53, CD63, CD81, CD82, TSG101J Cell Biol 200:373 (2013)
CD59, CD63, CD133, TSG101 FASEB J 23:1858 (2009)

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Exosomes can be isolated by ultracentrifugation or density gradient separation in addition to a precipitation approach. Exosomes can also be isolated by a magnetic approach using Dynabeads magnetic beads targeting exosome markers such as Human CD9, CD63, CD81, EpCAM or secondary antibody-coated Dynabeads magnetic beads using different antibodies against other exosomal surface markers.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

A range of different functions have been reported such as antigen presentation, apoptosis, angiogenesis, inflammation, and coagulation by protein/lipid exchange or activation of a signaling pathway. Exosomes provide a novel vehicle for genetic exchange between cells and mediate cell to cell communication. Exosomes also transport and propagate of infectious cargoes such as prion and retrovirus.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Exosomes are small, membrane-bound ovoid to cup shaped particles around 30-150 nm in size containing mRNA, microRNA, proteins, and lipids. Exosomes are released by normal, abnormal, or neoplastic cells into body fluid such as blood, urine, saliva, and breast milk. Exosomes originate from the endocytic compartment and are released from cells as multivesicular bodies (MVB) fused with plasma membrane (J Cell Biol 200:373 (2013)).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.