OverviewTake your EV insights to the next level by focusing on specific subpopulations“One of the biggest challenges for the EV field at this stage is addressing heterogeneity of secreted EVs, and heterogeneity within EV populations.”1Gain 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.Figure 1. Exo-Flow 2.0 beads have an outer silica coating which delivers undetectable background binding.“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 UniversityPowerful—analyze EV subpopulations for deeper insights into EV biology and biomarker discoveryFlexible—the basic kit extends the power of Exo-Flow 2.0 beads by giving you the ability to use the biotinylated antibody of your choice.Efficient—our Exo-Flow 2.0 beads can capture even small subpopulations of EVs to maximize your discoveriesLow-background—Exo-Flow 2.0 beads deliver undetectable background binding for targeted analysis of specific EV subpopulationsEasy-to-use—the magnetic bead-based workflow translates into quick and easy EV captureEach 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 AntibodyExo-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.#CD63EXOFLOW2-100A-SPEXOFLOW2-200A-TCCD9EXOFLOW2-105A-SPEXOFLOW2-205A-TCCD81EXOFLOW2-110A-SPEXOFLOW2-210A-TCTetraspanin Combo (CD63, CD9, and CD81)EXOFLOW2-150A-SPEXOFLOW2-250A-TCBasic Kit without antibodyEXOFLOW2-BASICA-SPEXOFLOW2-BASICA-TCReferencesWillms E, et al. Extracellular Vesicle Heterogeneity: Subpopulations, Isolation Techniques, and Diverse Functions in Cancer Progression. Front Immunol. 2018; 9: 738. PMCID: PMC5936763.How It WorksEasily analyze EV subpopulations with Exo-Flow 2.0 The Exo-Flow 2.0 workflow uses a simple, four-step workflow:Couple your biotinylated antibody to the magnetic streptavidin Exo-Flow 2.0 beadsUse the antibody-coupled magnetic beads to capture already-isolated EVs (compatible with ExoQuick®, SmartSEC™-HT, ultracentrifugation, or any other EV isolation method)Wash away unbound EVsPrep for analysisFor flow cytometry—stain with the included Red or Green DyeFor western blotting—resuspend beads in M-PER/RIPA lysis buffer (not included), add sample loading buffer load onto gelFor RNA analysis—resuspend beads in RNA lysis buffer of your choice and processFigure 2. The simple Exo-Flow 2.0 Workflow.Supporting DataSee the performance that Exo-Flow 2.0 technology deliversExo-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.Figure 3. Exo-Flow 2.0 delivers undetectable background binding when analyzed by western blot.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.Figure 4. Exo-Flow 2.0 delivers undetectable background binding when analyzed using flow cytometry.Understand surface marker differences between EV subpopulations with Exo-Flow 2.0We 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.Figure 5. Exo-Flow 2.0 supports flow cytometric analysis of EV subpopulations.Use Exo-Flow 2.0 to understand differences in cargo, such as miRNA levels, between EV subpopulationsWe 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.Figure 6. Use Exo-Flow 2.0 to understand differences in cargo, such as miRNA levels, between EV subpopulations.ResourcesMANUAL: Exo-Flow 2.0™ for Serum or PlasmaBrochure: Exosome Research Products and Services Related productsAnti-ALIX Antibody with Goat Anti-Rabbit HRP Secondary AntibodyShop NowAnti-EPCAM Antibody with Goat Anti-Rabbit HRP Secondary AntibodyShop NowEV-Entry SystemShop NowExosome Cyto-Tracer, pCT-CD81-RFPShop NowExosome Cyto-Tracer, pCT-CD9-RFPShop NowAnti-HSP70 Antibody with Goat Anti-Rabbit HRP Secondary AntibodyShop Now

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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

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ExoQuick and ExoQuick-TC Products

Product	Size	Catalog #
ExoQuick serum exosome precipitation solution (5 ml)	75 reactions	EXOQ5A-1
ExoQuick Plasma prep and Exosome precipitation kit (5 ml ExoQuick plus 500 ul Thrombin at 500U/mL), replaces EXOQ5TD-1 product	75 reactions	EXOQ5TM-1
Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL), replaces TDEXO-1 product	100 reactions	TMEXO-1
ExoQuick serum exosome precipitation solution (20 ml)	300 reactions	EXOQ20A-1
ExoQuick-TC for Tissue Culture Media and Urine (10ml)	10 reactions	EXOTC10A-1
ExoQuick-TC for Tissue Culture Media and Urine (50ml)	50 reactions	EXOTC50A-1
Exosome-depleted FBS Media Supplement - USA Certified	50 ml	EXO-FBS-50A-1
Exosome-depleted FBS Media Supplement - USA Certified	250 ml	EXO-FBS-250A-1
ExoQuick-LP Lipoprotein Pre-Clear & Exosome Precipitation Kit	5 reactions	EXOLP5A-1