Red Blood Cells
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Select “Plasma/Serum” for direct measurement in unfractionated complex biofluids like plasma or serum. Select “All Others” for cell culture supernatants and most other biofluids (eg. urine, CSF, saliva).
Current MISEV guidelines recommend measuring an EV associated protein. Although tetraspanin (TS) expression on EVs is heterogenous and not ubiquitous, the majority of EVs express some level of at least one of the common tetraspanins CD9, CD63, or CD81. The tetraspanin vTag™ cocktail labels these proteins and is sufficient comply with MISEV guidelines for most EVs.
Use a PE anti-TS vTag™ cocktail when sizing and counting EVs. If also using the kit to support no-wash immunofluorescent EV cargo measurement, you may select PE or another conjugate to adhere to principles of proper multicolor panel design. For example, a PE-Cy7 anti-TS cocktail is useful in some panels because it frees up the PE channel on the instrument and cargo of interest can then be measured using a PE conjugated antibody which are more widely available.
Cellarcus offers several bright vTag™ antibodies validated for sensitive, no-wash cargo detection using vFC™. Select other or use the site search to find available products. If you don’t see what you’re looking for, contact us. Our vTag™ catalog is always expanding.
NOTE: Lipo100™ is a synthetic vesicle size standard that enables size calibration when running a compatible assay such as vFC™. A standard is not an assay and simply incorporating a standard such as Lipo100™ is not sufficient to ensure a reliable robust method to characterizing EVs. It is, however, part of an assay that incorporates protocols, Lipo100™ and additional controls and calibrators, reagents, and analysis methods. A well-designed assay is required to satisfy recently published guidelines for EV characterization. For more info, learn about our vFC™ assay kit or contact us.
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Our standards are counted and characterized using a proprietary assay that specifically and accurately counts, sizes, and phenotypes vesicles (learn more). The data is cross-validated with widely used approaches such as Nanotracking Analysis (NTA) and Resistive Pulse Sensing (RPS). NTA and RPS count all particles (aggregates, cell debris, other non-vesicular material) and as such provide inaccurate, often inflated, vesicle concentrations. These technologies are inherently limited in their ability to specifically count vesicles. Standards sold by other manufacturers are limited by these technologies and sold in ug quantities with estimates of vesicle numbers. Our standards do not need to be sold as vesicle estimates based on total µg of protein. All of our standards are packaged based on our proprietary, highly accurate and reproducible vesicle counts. They are titrated into an optimal concentration for common assays expressed in vesicles per µl. The number of tests is based on the amount of standard required for a flow cytometric approach to vesicle analysis. Other techniques may require additional sample volume which will reduce the number of tests per vial. |
