Following stimulation, samples were stained with fluorescein isothiocyanate (FITC)-conjugated annexin V (AnV) (eBioscience, ThermoFisher, United Kingdom), to detect exposed PS (FL1), unless otherwise indicated, and PE-Cy7-conjugated anti-CD41 antibody (eBioscience, ThermoFisher), to distinguish plateletderived events. Samples were analyzed using a BD Accuri C6 flow cytometer. PE-Cy7 fluorescence (FL3) was used to trigger event acquisition. PS-positive platelet-derived EVs were defined as CD41+/AnV-positive (AnV+) events that were smaller than 1 μm. The 1 μm gate was set in forward scatter using 1 μm silica beads.19 (link) To monitor mitochondrial membrane potential, washed platelets (5×107/mL) were incubated with trimetylrhodamine methyl ester (TMRM; 100 μM, 30°C, 30 minutes; ThermoFisher) prior to treatment as indicated in the main text. TMRM fluorescence was acquired on FL2.
Fitc conjugated annexin 5 anv
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
FITC)-conjugated annexin V (AnV) is a fluorescent conjugate of the protein annexin V, which binds to phosphatidylserine (PS) exposed on the surface of apoptotic cells. This conjugate can be used to detect and quantify apoptosis in various cell types.
Automatically generated - may contain errors
2 protocols using fitc conjugated annexin 5 anv
1
Platelet-Derived Extracellular Vesicle Analysis
2
Platelet-Derived Extracellular Vesicle Analysis
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Following stimulation, samples were stained with fluorescein isothiocyanate (FITC)-conjugated annexin V (AnV) (eBioscience, ThermoFisher, United Kingdom), to detect exposed PS (FL1), unless otherwise indicated, and PE-Cy7-conjugated anti-CD41 antibody (eBioscience, ThermoFisher), to distinguish plateletderived events. Samples were analyzed using a BD Accuri C6 flow cytometer. PE-Cy7 fluorescence (FL3) was used to trigger event acquisition. PS-positive platelet-derived EVs were defined as CD41+/AnV-positive (AnV+) events that were smaller than 1 μm. The 1 μm gate was set in forward scatter using 1 μm silica beads.19 (link) To monitor mitochondrial membrane potential, washed platelets (5×107/mL) were incubated with trimetylrhodamine methyl ester (TMRM; 100 μM, 30°C, 30 minutes; ThermoFisher) prior to treatment as indicated in the main text. TMRM fluorescence was acquired on FL2.
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