Anti human hla dr apc ln3

For flow cytometric analysis, 5 × 10⁵ moDCs/mL were treated with 10 µg/mL BPE and 2.1 µg/mL Bet v 1 + 1.0 ng/mL LPS respectively (Fig. S1e). After incubation for 24 h, cells were washed with ice-cold DPBS and stained in 100 μl DPBS containing fluorescent-labelled antibodies for 30 min under protection from light. After extensive washing, staining was fixed by adding 4% (v/v) paraformaldehyde (PFA, Sigma-Aldrich) for 10 min. Then, cells were admitted into 200 µL DPBS containing 3 mM EDTA. The median fluorescence intensity (MFI) of 1 × 10⁴ cells was recorded for every sample using the FACS Canto™ II flow cytometer operated with FACS Diva Software v8.0 (BD Biosciences, Vienna, Austria). Data analysis was performed using the FlowJo^® Software v10.3 (FlowJo LLC, Ashland, Oregon, USA).
EBioscience™ fixable Viability Dye eFluor® 506, anti-human CD3 eFluor® 506 (clone: UCHT1), anti-human CD19 eFluor® 506 (HIB19), anti-human CD40 FITC (5C3), anti-human CD86 PE (IT2.2), and anti-human HLA-DR APC (LN3) were purchased from Thermo Fisher Scientific. Brilliant Violet 421™ anti-human CD1a (Hl149) was obtained from BioLegend (San Diego, CA, USA). APC-H7 mouse anti-human CD80 (L307.4) and PE-Cy™7 mouse anti-human CD83 (HB15e) were purchased from BD Biosciences.