The informed consent and related protocols were approved by Emory University institutional review boards. Citrated whole blood was obtained from healthy volunteers, from which platelet-rich plasma (PRP) was prepared [30 (link)]. Platelet counts were measured using a Sysmex XP-300 analyzer (Lincolnshire, IL). To measure platelet binding, PRP was incubated with 60 nM noted protein for 10 minutes at room temperature. After washing the samples were mixed with 1 µg/mL FITC-labeled anti-His-tag MAb, fixed and analyzed on a BD FACS Canto-II instrument using FlowJo. The signal was quantitated by the median fluorescence intensity (MFI) for the entire cell population (10,000 cells).
Protocol detail
Platelet Protein Binding Assay
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Blood
Cells
Fitc
Fluorescence
Healthy volunteers
Institutional review boards
Platelet
Platelet counts
Platelet rich plasma
Protein
Corresponding Organization : Emory University
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Variable analysis
independent variables
- Protein (60 nM)
- Platelet binding (measured by median fluorescence intensity (MFI))
- Incubation time (10 minutes)
- Temperature (room temperature)
- Platelet-rich plasma (PRP) obtained from healthy volunteers
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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