“One of the biggest challenges for the EV field at this stage is addressing heterogeneity of secreted EVs, and heterogeneity within EV populations.”1
Gain powerful insights into extracellular vesicle (EV) biology and analyze EV subpopulations for therapeutics development, biomarker discovery, and more with SBI’s Exo-Flow™ 2.0 Basic Kit without antibody (Streptavidin beads + reagents) – for Serum or Plasma. With Exo-Flow 2.0 technology, we’ve re-engineered our original Exo-Flow beads (Figure 1) to deliver virtually undetectable background binding. The result is highly specific capture of EVs expressing the surface marker of your choice using biotinylated antibody that you supply, so you can easily analyze specific EV subpopulations by flow cytometry, western blot, or other downstream technology. Take your EV insights to the next level with Exo-Flow 2.0.
“The new Exo-Flow 2.0 beads are much easier to see during the processing steps (sample capturing, washing, staining, etc.) which prevents accidental loss of the sample. I am also able to acquire more events with the flow cytometer with the new Exo-Flow 2.0 beads.”
—Ananthu Pucha, DeKalb Veterans Affairs Hospital at Emory University
Each Exo-Flow 2.0 Basic Kit without antibody (Streptavidin beads + reagents) – for Serum or Plasma comes with enough reagents for 30 reactions of 150 – 300 µg protein equivalent of EVs (the actual amount of input protein equivalent depends on your EV isolation method).
Our Exo-Flow 2.0 technology is available as both individual tetraspanin (CD63, CD9, and CD81) and combination kits, as well as a basic kit that can be used with the biotinylated antibody of your choice. Note that unlike the original Exo-Flow technology, Exo-Flow 2.0 kits are specific for isolating EVs from either serum/plasma or tissue culture medium, so be sure to choose the right kit for your needs.
See all available Exo-Flow 2.0 kits in the table below.
Biotinylated Antibody | Exo-Flow 2.0 Kit for EV Isolation from Serum or Plasma Cat.# | Exo-Flow 2.0 Kit for EV Isolation from Tissue Culture Medium Cat.# |
---|---|---|
CD63 | EXOFLOW2-100A-SP | EXOFLOW2-200A-TC |
CD9 | EXOFLOW2-105A-SP | EXOFLOW2-205A-TC |
CD81 | EXOFLOW2-110A-SP | EXOFLOW2-210A-TC |
Tetraspanin Combo (CD63, CD9, and CD81) | EXOFLOW2-150A-SP | EXOFLOW2-250A-TC |
Basic Kit without antibody | EXOFLOW2-BASICA-SP | EXOFLOW2-BASICA-TC |
References
The Exo-Flow 2.0 workflow uses a simple, four-step workflow:
Exo-Flow 2.0 delivers undetectable background binding when analyzed by western blot
We isolated EVs from HEK293 cells and serum using ExoQuick®-TC and ExoQuick® Exosome Isolation Reagents, captured specific EV subpopulations using Exo-Flow 2.0 beads coated with either anti-CD63, anti-CD9, anti-CD81, or a no-antibody control, and analyzed the captured EVs by western blot probing for the exosome-specific marker TSG101 (Figure 3). The absence of detectable signal in the no-antibody control lane demonstrates the low background binding of Exo-Flow 2.0 technology. The varying signal intensity seen in Figure 3 also highlights the heterogeneity of EV subpopulations.
Exo-Flow 2.0 delivers undetectable background binding when analyzed using flow cytometry
We isolated EVs from HEK293 cells using ExoQuick-TC and captured the subpopulation of EVs expressing CD9 (Figure 4A, dark red trace) and the subpopulation of EVs bearing CD63 (Figure 4A orange trace). We also isolated EVs from serum using the SmartSEC HT EV Isolation system and captured the subpopulation of EVs bearing CD81 (Figure 4B). After staining bead-bound EVs and washing away excess stain, we analyzed the EV subpopulations by flow cytometry. In both plots, distinct subpopulations of EVs can be seen—in Figure 4A, the peak seen on the red trace indicates a subpopulation of EVs bearing CD9 and the separate peak seen on the orange trace indicates a subpopulation of EVs bearing CD63. In Figure 4B, the peak seen on the green trace indicates a subpopulation of EVs bearing CD81.
In both plots, the overlap of signal in the beads-only and no-antibody traces demonstrates the low background binding of Exo-Flow 2.0 technology.
Understand surface marker differences between EV subpopulations with Exo-Flow 2.0
We isolated EVs from serum using SmartSEC HT, captured subpopulations of EVs using Exo-Flow 2.0 beads coupled with either biotinylated CD9 (Figure 5A) or CD63 (Figure 5B) antibodies, and analyzed these subpopulations for the presence of CD14, a myeloid lineage marker, using flow cytometry (Figure 5). The data show that EVs captured using anti-CD9 Exo-Flow 2.0 beads are more likely to also bear CD14 than EVs captured using anti-CD63- Exo-Flow 2.0 beads, demonstrating the effectiveness of Exo-Flow 2.0 technology for EV subpopulation analysis and enabling better biomarker screening.
Use Exo-Flow 2.0 to understand differences in cargo, such as miRNA levels, between EV subpopulations
We isolated EVs from HEK293-conditioned medium using ExoQuick-TC, captured subpopulations of EVs using Exo-Flow 2.0 beads coupled with either biotinylated CD63 or CD81 antibodies, and analyzed these subpopulations for the presence of miR-122 (Figure 6). The data show a statistically significant difference in the amount of miR-122 present in these EV subpopulations, with ~3.6-fold more miR-122 found in CD81 EVs than CD63 EVs, demonstrating the effectiveness of Exo-Flow 2.0 technology for EV subpopulation analysis.
System Biosciences,简称SBI,美国加州湾区新成立的生技公司,致力于独特、创新生物技术之开发,以研发利于基因及蛋白质功能鉴定、研究之崭新方法和工具为宗旨。 现阶段研发重心为RNA干扰(RNAi)研究之相关工具。 System Biosciences (SBI) 致力于开发独特、革新的技术,为客户研究蛋白组学和基因组学功能提供研究工具。SBI 是专业的慢病毒产品公司,提供基于慢病毒的所有相关产品、质粒、试剂盒及相关配套试剂和慢病毒延伸产品如IPS细胞多功能性诱导试剂盒和RNAi筛选文库。
SBI focuses on developing unique, innovative technologies that provide researchers with the tools to investigate and understand genomic and proteomic function. Our mission is to provide tools for the genome-wide analysis of the mechanisms that regulate cellular processes and biological responses.
Headquartered in Palo Alto about 30 miles south of San Francisco, SBI is geographically surrounded by highly successful research biotech companies (e.g., Affymetrix, Life Technologies, and Genentech) and some of the premier life science institutes in the world, including Stanford and the University of California, San Francisco.
As a small private biotechnology company, SBI capitalizes on a rich network of consultants and colleagues and continually collaborates to accelerate its development of innovative and novel applications. Currently, SBI has partnerships with over thirty scientists from several institutes.System Biosciences (SBI) consists of a highly motivated team committed to realizing the potential of its mission to develop and bring to market unique and innovative technology to investigate and understand genomic and proteomic function.
Possessing a diverse range of experience and knowledge, SBI's management and staff bring the skills, talent, and interest needed to support the company's continuing rapid growth.
A strong and proven background in creative research and product development with proficiency in genetic analysis, microarray technology, and cell biology
Extensive business development and management experience
A keen understanding of the needs of life scientists
Expertise in developing, marketing, and supporting consistent, high-quality research products
A clear focus on the need to build and maintain lasting customer relationships through support, service, and close customer interaction
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