The Effects of Shear Stress on Platelets Treated with Antibodies Against Mechanosensory Receptors
Published: January 31, 2024
Abstract
Source: Quach, M. E. et al., A Uniform Shear Assay for Human Platelet and Cell Surface Receptors via Cone-plate Viscometry. J. Vis. Exp. (2019)
This video demonstrates the method to assess the impact of shear stress on platelets. Antibody treatment induces dissociation and conformational changes in mechanoreceptor complexes, activating the platelets and expressing markers on the surface. These markers are observed via immunostaining and flow cytometry.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Blood Draw and Platelet Isolation Draw human blood from consenting healthy adult donors via venipuncture on the day of the experiment into 3.8% trisodium citrate. One 4.5 mL tube of blood is sufficient to yield enough platelet-rich plasma (PRP) for 20-25 conditions in donors whose platelet counts are close to 250 x 103 per μL…
Disclosures
The authors have nothing to disclose.
Materials
| APC anti-human CD62P (P-Selectin) | BioLegend | 304910 | |
| Brookfield Cap 2000+ Viscometer | Brookfield | – | |
| FITC-conjugated Erythrina cristagalli lectin (ECL) | Vector Labs | FL-1141 | |
| Pooled Normal Human Plasma | Precision Biologic | CCN-10 | |
| Vacutainer Light Blue Blood Collection Tube (Sodium Citrate) | BD | 369714 | |
| Vacutainer Blood Collection Set, 21G x ¾" Needle | BD | 367287 |
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Cite This Article
The Effects of Shear Stress on Platelets Treated with Antibodies Against Mechanosensory Receptors. J. Vis. Exp. (Pending Publication), e21930, doi: (2024).