Anti cd11c apc

After thawing, PBMCs (N = 74) were analysed by flow cytometry using anti-CD11b-PE, anti-CD14-PerCP, and anti-CD11c-APC (Immunotools, Friesoythe, Germany), anti-CD33-APC, and anti-HLA-DR-APCH7 (BD Biosciences, San Jose, CA, USA). Viability was assessed by flow cytometry using the LIVE/DEAD TM Fixable Violet Dead Cell Stain Kit (Thermo Fisher Scientific, Vienna, Austria). MDSC were identified by combining anti-CD14, anti-HLA-DR, anti-CD33, and anti-CD11b. Data analysis was performed using FlowJo (v.10, Ashland, OR, USA). The percentage of cells and the count of cells/µL were determined.

The expression of CD11b and CD11c was measured by flow cytometry at different time points during the LPS induced cell activation. LPS treated and untreated control cells were harvested after 30 minutes, 24 hours or 48 hours, and washed with ice cold PBS (phosphate buffered saline solution) supplemented with 0.4% sodium azide and 1% FCS. Cells were labelled with anti-CD11b-FITC and anti-CD11c-APC (ImmunoTools GmbH) for 20 minutes on ice. The expression of the receptors was compared to the appropriate untreated control sample at each time point. Samples were analyzed on Cytoflex flow cytometer (Beckman Coulter) using CytExpert software for data acquisition and Kaluza Analysis software for data analysis.