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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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vCal™ MESF Calibrator nanoBeads are surface dyed beads calibrated against fluorescence of solutions of known concentration of the same fluorophore. Each product contains 3 populations of beads with increasing amounts of the specified fluorophore. MESF beads are necessary to achieve quantitative immunofluorescence of EV surface cargo which should be performed to ensure EV cargo data is comparable across time and labs and also to understand limit of detection for an instrument in a particular channel to interpret negative events when running samples. The 800nm size is suitable for detection on flow cytometers with lower scatter sensitivity allowing assessment of an instrument’s sensitivity in a given channel independent of scatter performance. Fluorescence intensities these beads are in a relevant range for vesicle immunostaining and consequently much dimmer than other commercial products.

Fluorophore : PE. Others coming soon.
Bead Population(s): 3 populations. See COA for MESF units calculated for each population.
Concentration: 1×107 beads per ml.
Bead Size: 800nm
Buffer: 0.1um filtered PBS +0.09% sodium azide.
Storage Conditions: Store at 4C. Minimize temperature cycling when handling (e.g. when using beads, keep the tube in the refrigerator as much as possible).

25 ul (5 tests)

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