Red Blood Cells
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Select a tetraspanin marker to comply with MISEV guidelines which recommend confirming the presence of an EV associated protein. Tetraspanins in the antibody cocktail are heterogeneously expressed, but present on most EVs (see example). Use the PE cocktail when sizing and counting EVs. When also using the kit to support no-wash immunofluorescent EV cargo measurement, select another conjugate to enable proper panel design. For instance, PE-Cy7 is useful beause it frees up the PE channel and other good channels on the instrument that detect bright, widely available antibody conjugates to enable sensitive quantitation of experimentally important proteins (including our vTag™ mAbs, validated for vFC™, searchable below).
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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. |
