Blood count analysis
anti-CD15-PacificBlue
FLUOR, LYSIS
Antibodies for blood count analysis: multi-color flow cytometry was performed using Beckman Coulter’s custom design service and its dry coating technology, and custom tubes containing anti-CD16-FITC (clone 3G8), anti-CD56-PE (clone N901), anti-CD19-PE-Cy5.5 (clone J3-119), anti-CD14-PE-Cy7 (clone RMO52), anti-CD4-APC (clone 13B8.2), anti-CD8-Alexa Fluor 700 (clone B9.11), anti-CD3-APC-Alexa Fluor 750 (clone UCTH1), anti-CD15-PacificBlue (clone 80H5) and anti-CD45-KromeOrange (clone J.33) were produced.Staining protocol: 100 μl of total heparinized blood was added to a DURAClone tube, vortexed immediately for 15 s and incubated for 15 min at room temperature in the dark. Two milliliters of red blood lysis solution (VersaLyse solution, A09777, Beckman Coulter) containing 50 μl of the fixative agent IOTest 3 Fixative Solution (A07800, Beckman Coulter) was added, inverted and incubated for 15 min in the dark. Then, 100 µl of counting beads (Flow-Count Fluorospheres, 7547053, Beckman Coulter) was added before acquisition on a Canto II cytometer (BD Biosciences).