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2 protocols using ltc4 enzyme linked immunoassay eia kit

1

Mast Cell Signaling and Inflammation Assays

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RPMI1640, fetal bovine serum (FBS) and the enhanced chemiluminescence (ECL) Western blot detection reagent were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, United States). Mouse anti-dinitrophenyl (DNP) IgE was purchased from Sigma Chemicals (St. Louis, MO, United States). DNP-human serum albumin (HSA) was from Biosearch Technologies (Petaluma, CA, United States). The antibodies specific for phospho-ERK1/2, ERK1/2, phospho-p38, p38, phospho-JNK1/2, JNK1/2, phospho-PLCγ, phospho-IκBα, IκBα, phospho-IKKα/β, β-actin, and the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, United States). The antibodies specific for phospho-cPLA2, NF-κB p65, lamin B, LAT, Lyn, Fyn, and Syk, as well as Bay 61-3606 reagent were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, United States). The LTC4 enzyme linked immunoassay (EIA) kit, and the antibody for COX-2 were from Cayman Chemical (Ann Arbor, MI, United States). Histamine ELISA kit was purchased from Demeditec Diagnostics GmbH (Kiel, Germany).
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2

Mast Cell Activation Assay

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RPMI1640, fetal bovine serum (FBS), and the enhanced chemiluminescence (ECL) Western blot detection reagent were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Mouse anti-dinitrophenyl (DNP) IgE was purchased from Sigma Chemicals (St. Louis, MO, USA). DNP-HSA was from Biosearch Technologies (Petaluma, CA, USA). The antibodies specific for phospho-ERK1/2, ERK1/2, phospho-p38, p38, phospho-PLCγ, phospho-IκB, IκB, phospho-IKKα/β, β-actin, and the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). The antibodies specific for phospho-cPLA2, NF-κB p65, lamin B, Lyn, Fyn, and Syk, as well as Bay 61-3606 reagent were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The LTC4 enzyme linked immunoassay (EIA) kit, and the antibody for COX-2 were from Cayman Chemical (Ann Arbor, MI, USA). Histamine enzyme linked immunosorbent assay (ELISA) kit was purchased from Demeditec Diagnostics GmbH (Kiel, Germany). AB23A was obtained from Push Bio-technology Co., Ltd (Chengdu, China), of which the purity is ≥98%.
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