Anti human cd117 pe

Flow cytometry analyses were performed using either an LSR Model II BD FACSArray (BD Biosciences, Oxford, UK) or a Novocyte (Acea Biosciences, San Diego, CA, USA) flow cytometer. Cell-sorting experiments were performed using either Influx or FACSAria fluorescence-activated cell sorters (both from BD Biosciences). Antibodies used were anti-human CD11b-PE, anti-human CD14-FITC (fluorescein isothiocyanate), anti-human CD86-PerCP-eFluor710, and anti-Human CD117-PE (eBioscience, Hatfield, UK).

Dulbecco's Modified Eagle's Medium (DMEM), Iscove’s modified Dulbecco’s medium (IMDM), fetal bovine serum (FBS) and penicillin-streptomycin solution were purchased from Life Technologies (Gibco). L-glutamine, ascorbic acid, 1-thioglycerol, HEPES buffer, nonessential amino acids and sodium pyruvate were purchased from Sigma. Human stem cell factor (SCF), interleukin 6 (IL-6) and interleukin 3 (IL-3) were obtained from Peprotech. Recombinant Human MIF (Z03159-50) was obtained from Genscript. 6-well plates and 10-cm ultra-low-attachment dishes were purchased from Corning. Anti-chymase (MA5-11717) and anti-tryptase (MA5-11711) antibodies were purchased from Thermo Scientific Pierce Antibodies. Human IgE, anti-IgE antibody (ab75673) and anti-MIF antibody (ab7207) were bought from Abcam. Phycoerytrin (PE)-labeled anti-CD34 antibody (HPCA-2) and PE-IgG isotype control were purchased from Becton Dickinson (BD). Anti-human FcεRI-FITC and anti-human CD117-PE were purchased from eBioscience. SPI Bio histamine EIA kit was obtained from Bertin Pharma. [3H]-proline was from Perkin-Elmer.