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Purification and Activation of Platelets from Murine Whole Blood
Overview
In this video, we describe a protocol for purifying platelets from murine whole blood and their subsequent activation with the platelet agonist thrombin. When analyzed with flow cytometry, the purified resting platelets are characterized based on the expression of their cell-surface marker CD41, whereas purified activated platelets co-express both the surface markers CD41 and CD62P.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Platelet Purification
NOTE: Platelet purification should be carried out at room temperature to prevent their degradation. Make sure that the temperature of the reagents and instruments is between 18 to 22 °C. To prevent platelet degradation, fast pipetting, and vigorous shaking should.......
Representative Results
Figure 1. Mouse blood before and after centrifugation with Iohexol gradient medium. (A) Iohexol and blood form two separate layers before centrifugation if the blood sample is added carefully. Left panel shows schematic diagram and right panel shows the actual image. (B) Four separate layers are seen after centrifugation of whole blood with gradient medium. Left panel shows schematic diagram of .......
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Source: Narciso, M. G., et al. Purification of Platelets from Mouse Blood. J. Vis. Exp. (2019).
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