American Society of Hematology

Figure 4

$Figure 4. αTAT ELISA analyses of reconstituted PRP. WB was drawn via phlebotomy into 0.1 mg/mL of CTI and subjected to differential centrifugation to obtain WB, PRP, PPP, buffy coat, and packed RBCs. An aliquot of CTI blood was “mock fractionated” via centrifugation at 150g, followed by remixing before activation by Tf. (A) αTAT ELISA analyses were performed on a time course from a representative subject's WB (○) and mock fractionated blood (●) that was subjected to 5pM relipidated rTf. Samples were quenched at various intervals with EDTA and Phe-Pro-Arg chloromethylketone and analyzed via αTAT ELISA. (B) CTI blood was subjected to differential centrifugation to obtain WB, PRP, PPP, buffy coat (WBC surrogate), and packed RBCs. The latter 3 fractions were used to reconstitute PRP to each subject's physiologic platelet concentration. αTAT ELISA analyses were performed on blood fraction time courses from 5 subjects WB (○), PRP + PPP (■), PRP + buffy coat (♢), and PRP + packed RBCs (▾) subjected to 5pM relipidated rTf. Samples were quenched at various intervals with EDTA and Phe-Pro-Arg chloromethylketone and analyzed via αTAT ELISA. Data are shown as the means ± SEM (n = 5).$

αTAT ELISA analyses of reconstituted PRP. WB was drawn via phlebotomy into 0.1 mg/mL of CTI and subjected to differential centrifugation to obtain WB, PRP, PPP, buffy coat, and packed RBCs. An aliquot of CTI blood was “mock fractionated” via centrifugation at 150g, followed by remixing before activation by Tf. (A) αTAT ELISA analyses were performed on a time course from a representative subject's WB (○) and mock fractionated blood (●) that was subjected to 5pM relipidated rTf. Samples were quenched at various intervals with EDTA and Phe-Pro-Arg chloromethylketone and analyzed via αTAT ELISA. (B) CTI blood was subjected to differential centrifugation to obtain WB, PRP, PPP, buffy coat (WBC surrogate), and packed RBCs. The latter 3 fractions were used to reconstitute PRP to each subject's physiologic platelet concentration. αTAT ELISA analyses were performed on blood fraction time courses from 5 subjects WB (○), PRP + PPP (■), PRP + buffy coat (♢), and PRP + packed RBCs (▾) subjected to 5pM relipidated rTf. Samples were quenched at various intervals with EDTA and Phe-Pro-Arg chloromethylketone and analyzed via αTAT ELISA. Data are shown as the means ± SEM (n = 5).

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