Exosome LabelingExoGlow-Vivo EV Labeling Kit (Near IR) for in vivo imaging of EVs in animal models
• Optimized for in vivo imaging of EVs in animal models
• NIR spectrum enables deep tissue illumination
• Eliminates background from auto-fluorescence
• Non-lipophilic dye overcomes background from non-specific labeling
• From EV isolation to injection-ready labeled EVs in less than 1 hour
• Ideal for in vivo EV tracking, biodistribution, and kinetic studies
• Follow EVs in organ homing studies
• Allows ADME studies of EVs as delivery vehicles
• Compatible with all tested EV isolation methods (incl. ExoQuick, ultracentrifugation, column-based)
• Usable with as little as 250 µg of EVs
Visualizing extracellular vesicles (EVs) in cells and organisms has been technically difficult due to the high background from auto-fluorescence in the UV and visible areas of the spectrum and a lack of specificity from the typically lipophilic dyes. SBI’s ExoGlow-Vivo EV Labeling Kit (Near IR) is the first reagent specifically designed to overcome these problems through the use of a proprietary, non-lipophilic dye that emits in the near infrared (NIR) range (excitation at 784 nm; emission at 806 nm). Delivering a level of specificity and sensitivity that takes the guesswork out of tracking EVs in vivo, ExoGlow-Vivo is ideal for EV biodistribution and kinetic studies needed to fully realize the value of EVs in basic research and translational applications.
ExoGlow-Vivo dye delivers minimal background.
A. Human mesenchymal stem cell-derived EVs were labeled with ExoGlow-Vivo dye and unbound dye removed via ultracentrifugation and a wash. EVs were administered intravenously via the superficial temporal vein into post-natal day-4 FVB mice. Animals were imaged at various time points using an IVIS® In Vivo Imaging System (PerkinElmer). Control refers to supernatant from wash step (i.e. free dye). B. Dissection after 24-hours shows the preferential accumulation of labeled EVs in specific organs and very low residual background signal from free dye. Data courtesy of Gareth Willis, PhD., Harvard Medical School and Boston Children’s Hospital.
ExoFlow-ONE EV Labeling Kits for Flow Cytometry
• Directly detect EVs by specifically labeling internal EV components
• Accurately estimate the size of labeled EVs
• Ensure proper run-to-run calibration with the included size standards
• Learn more from a single sample by multiplexing with our range of spectrally separated ExoFlow-ONE dyes
• Achieve near single-vesicle visualization* with our proprietary ExoFlow-ONE dyes that deliver
>High quantum efficiency
>Negligible background signal when not bound to EVs
Harness the power of flow cytometry for EV analysis
Finally, you can take full advantage of the power of flow-based methods for analyzing extracellular vesicles (EVs) with SBI’s exclusive ExoFlow-ONE Gemstone dyes. By specifically labeling internal EV components with one of these proprietary, high quantum efficiency dyes, you can achieve near single-vesicle resolution* for more powerful flow cytometry and FACS studies, and uncover greater insights into EV biology.
The ExoFlow-ONE Ruby Red Gemstone Dye Kit comes with enough labeling reagents for 25 reactions**. Each kit also includes 1.5 ml of silica bead size standards (110 nm – 1300 nm), two of which are fluorescent. The silica beads, sourced from Apogee Flow Systems, are specifically designed to produce a refractive index and light scattering patterns similar to biological particles, enabling more accurate estimation of EV particle size.
*Requires a specialized flow cytometry imager capable of high-resolution vesicle analysis (e.g. Amnis® ImageStreamX Mark II, Apogee Micro-PLUS).
**Each reaction is defined as 1 ul of working dye combined with 200 – 500 ug of EV protein resuspended in 500 ul of buffer.
Choose the right ExoFlow-ONE dye for your project
• Monitor exosome trafficking in real-time
• Stable lentivector-based system for tracing
• Bright and photostable GFP/RFP is non-disruptive to cells
Molecular trafficking is a dynamic process in eukaryotic cells and the Cyto-Tracers provide the ability to light up cell compartments to monitor movement and localization of organelles and to trace endocytosis and exocytosis.
The tetraspanin proteins CD63, CD9 and CD81 are common biomarkers for exosomes.
Lentivector-based Cyto-Tracers expressing tetraspanin CD63, CD9 or CD81 fused to GFP or RFP mark exosomes and enable more long-term and more in-depth experimentation. The exosomal Cyto-Tracers can be used in transfections as well as packaged into virus to create stable GFP/RFP tracer cell lines in primary cells, tumor cell lines and stem cells. These cell lines secrete glowing exosomes which can be isolated using ExoQuick Exosome Precipitation Solution for downstream functional delivery and tracking studies.
Exosomes isolated using ExoQuick-TC Exosome Precipitation Solution can be transferred between cells.
A stable 293TN cell line overexpressing the exosomal Cyto-Tracer CD63-GFP fusion protein (Cat# CYTO120-PA-1-SBI) was generated. The medium from the cells was collected 48 h after plating and the exosomes from the media were precipitated using ExoQuick-TC Exosome Precipitation Solution. The exosome pellet recovered was resuspended in 30 μl PBS and 10 μl was added to newly plated HT1080 cells.
HT1080 cells were visualized 72 h after the addition of the CD63-GFP labeled exosomes and then re-plated. Following another 24h, the cells were again visualized for GFP fluorescence and imaged. The exosomes appear to dock with the cells within 72 h and some are found to be internalized after 96 h.
For the precipitation of the glowing exosomes use SBI´s ExoQuick/ExoQuick-TC Exosome Precipitation Solution for subsequent functional delivery and tracking studies, see link below.
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