Anti cd14 apc 61d3

For flow cytometry analysis, 50 μl of peripheral venous blood was used to obtain 100,000 leukocytes via the inertial microfluidic system (Fig. 1). Leukocytes were incubated for 20 min at RT with the following antibodies to human proteins, with clones noted in parentheses: anti-CD45 PerCP (HI30), anti-CD66b Pacific blue (G10F5), anti-CD16 APC-Cy7 (3G8), anti-CD69 FITC (FN50), pHrodo (PE), anti-CD62L Brilliant Violet 510 (DREG-56), anti-CD42b Alexa Flour 700 (HIP1) (all from BioLegend), and anti-CD14 APC (61D3) and anti-CD11b PE-Cy7 (ICRF44) (from Thermofisher). After staining, the cells were lysed and fixed with 2 ml of 1:4 dilution of Lyse/fix Buffer 5X (BD Phosflow) with dH₂O for 15 min at RT. Data were acquired from the BD LSR Fortessa flow cytometer and analyzed using Flowjo software version 10.1 (Tree star). PMN (CD45⁺SSC^HiFSC^Hi) subsets were determined by CD16 and CD66b surface expression. Monocyte (CD45⁺SSC^LowFSC^High) subsets were determined by CD14 and CD16 surface expression. Molecules of leukocyte activation were assessed, namely expression of CD62L, CD11b, CD69, and CD42b.