Human cd45 microbeads reagent

According to the findings of a previous study, density centrifugation and CD45+ cell depletion were used to isolate and purify platelets from whole blood.23 (link) Briefly, peripheral blood samples were treated with citrate dextrose to obtain the plasma, which comprised 85 mM trisodium citrate, 78 mM citric acid, and 111 mM glucose. The samples were then centrifuged at 80×g for 10 min at 22°C. To collect the platelets, the PRP was centrifuged at 1000 x g for ten minutes at a temperature of 22°C. after being mixed with 2 mM of ethylenediaminetetraacetic acid. Bead buffer (0.02% KCl, 0.8% NaCl, 0.024% KH2PO4, 0.144% Na2HPO4, 0.5% bovine serum albumin, 2 mM ethylenediaminetetraacetic acid) and 40 µL human CD45 MicroBeads reagent were added to the resuspended platelets in a volume of 3 mL. (Miltenyi Biotec, Germany). Using a MACS bead separation system (Miltenyi Biotec, Germany), we were able to isolate platelets free of leukocytes to greater than 99.99% after incubating them at room temperature for 45 min with gentle mixing. RNA from the platelets was isolated with the use of TRIzol (Life Technologies, USA). An Agilent Bioanalyzer was used throughout the process of quality control screening for each and every RNA sample.