Rabbit anti h4r

Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91^phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

Formalin fixed paraffin embedded human esophageal biopsy sections were rehydrated in sodium citrate buffer and blocked with Startingblock T20 blocking buffer (Thermo scientific, Rockford, IL). Sections were then incubated with rabbit anti-H1R (1:100, Santa Cruz, Santa Cruz, CA), goat anti-H2R (1:200, Santa Cruz), or rabbit anti-H4R (1:100, Santa Cruz) diluted in PBT overnight at 4°C. Mouse anti-MBP (1:100, Millipore Temecula, CA) was used to co-stain for eosinophils. After washing with PBS, Cy3 or Cy2-conjugated secondary antibody was applied for 2 hours at room temperature (1:100, Jackson Immunoresearch West Grove, PA). Cells were washed in PBS and stained with 4,6-diamidino-2-phenylindole (1:10,000, Invitrogen) according to manufacturer’s recommendations. Coverslips were applied after the application of mounting media (Biomeda, Foster city, CA), and sections were imaged using Olympus BX51 fluorescent microscope.

Dulbecco’s modified Eagle’s medium (DMEM) and fetal calf serum (FCS) were purchased from Gibco–BRL (Grand Island, NY, United States). Lipopolysaccharide (LPS, from Escherichia coli 0111:B4), disodium cromoglycate (cromolyn) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Fluorescenc mounting medium with DAPI and anti-mast cell Tryptase antibody were purchased from Abcam (HK, China). Rabbit anti-Iba1 antibody was purchased from Wako Chemicals USA, Inc. Rabbit anti-H₁R and anti-H₂R antibodies were purchased from Alomone Labs (Jerusalem, Israel). Rabbit anti-H₃R antibody was purchased from Abcam (Hong Kong, China). And rabbit anti-H₄R was purchased from Santa Cruz (CA, United States). Fluorescein isothiocyanate (FITC) – conjugated mouse anti-OX-42 antibody and isotype control, phycoerythrin (PE)-conjugated goat anti-rabbit secondary antibody were purchased from BD (BD Biosciences, United States). Cytokines (TNF-α, IL-6, IL-1β, INF-γ, RANTES, and IL-10) measured kit was purchased from MERCK Millipore Corporation (Billerica, MA, United States). Specific rabbit anti-p38, anti-Phospho-p38, anti-JNK, anti-Phospho-JNK, anti-ERK, anti-Phospho-ERK, anti-AKT, anti-Phospho-AKT, and anti-Phospho-NF-κB p65 antibodies, and goat anti-rabbit secondary antibody were obtained from Cell Signaling (Beverly, MA, United States).