Dnp human serum albumin has

RPMI-1640, Modified Eagle Medium (MEM), non-essential amino acid solution, and penicillin-streptomycin were acquired from Hyclone (Logan, Utah, USA). Imperatorin was purchased from ChromaDex (Irvine, CA, USA). Mouse anti-dinitrophenyl (DNP) IgE and DNP-human serum albumin (HAS) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The recombinant mouse SCF was purchased from STEMCELL Technologies Inc (Vancouver, BC, Canada). The primary antibodies used in the experiments were as follows: rabbit polyclonal antibodies specific for phospho-ERK1/2, ERK1/2, phosphop38, p38, phospho-JNK, JNK, phospho-PLCγ1, phospho-Akt, Akt, phospho-IκBα, IκBα, phospho-IKKα/β, and β-actin from Cell Signaling Technology, Inc. (Beverly, MA, USA); rabbit polyclonal antibodies for 5-LO, and phospho-cPLA₂ from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Specific inhibitors for NF-κB (pyrrolidine dithiocarbamate, PDTC) and MAPKs (SB203580, PD98059, and SP600125) were obtained from Sigma. The horseradish peroxidase-conjugated goat anti-rabbit secondary antibody was from Cell Signaling. 1×RIPA buffer, NE-PER Nuclear Protein Extraction kit, phosphatase/protease inhibitor cocktail, and enhanced chemiluminescence (ECL) detection reagent were from Pierce (Rockford, IL, USA). The enzyme immunoassay (EIA) kits for LTC₄ and PGD₂, and COX-2 antibody were purchased from Cayman Chemicals (Ann Arbor, MI, USA).