Sodium monoiodoacetate mia

The following commercial materials were used: Alexa Fluor 546 conjugated anti-rabbit IgG antibody and Alexa Fluor 488-conjugated anti-goat IgG antibodies were from Thermo Fisher Scientific (Yokohama, Japan); anti-mast cell tryptase (V-13) antibody against mMCP-6 was from Santa Cruz Biotechnology (Santa Cruz, CA); anti-PAR2 antibody was from Bioss Inc. (Woburn, MA); anti-NF-H antibody was from Gene Tex Inc. (Irvine, CA); anti-GFP antibody was from Novus Biologicals (Centennial, CO); PAR2 antagonist peptide (FSLLRY-NH₂) (TOCRIS), PAR2 agonist peptide (SLIGKV-NH₂) (TOCRIS) and ATP Detection Assay Kit (Cayman Chemicals) were from Funakoshi Co., Ltd. (Tokyo, Japan); WEHI-3 cells were from American Type Culture Collection; RNA extraction kit (Thermo Fisher Scientific) and real time PCR kit (TAKARA) were from Takara Bio Inc. (Kusatsu, Japan); tryptase substrate (S-2288) was from Chromogenix (Milano, Italy); ATP, sodium monoiodoacetate (MIA) and apyrase were from Sigma-Aldrich Japan (Tokyo, Japan).

KML29 (MAGL inhibitor; 1-piperidinecarboxylic acid, 4-[bis(1,3-benzodioxol-5-yl)hydroxymethyl]-, 2,2,2-trifluoro-1-(trifluoromethyl)ethyl ester) was obtained from Med Chem Express Ltd. (Monmouth Junction, NJ, USA). Celecoxib (CXB; COX-2 inhibitor; 4-[5-(4-methylphenyl0-3-(trifluoromthyl)-1H-pyrazol-1-yl]-benzenesulfonamide), AM281 (CB1 receptor antagonist; 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4-morpholinyl-1H-pyrazole-3-carboxamide), and AM630 (CB2 receptor antagonist; 6-iodo-2-methyl-1-(2-morpholin-4-ylethyl)indol-3-yl]-(4-methoxyphenyl)methanone) were obtained from Cayman Chemicals (Ann Arbor, MI, USA). Rhodamine 6G, cremophor, dimethyl sulphoxide (DMSO), urethane, and sodium monoiodoacetate (MIA) were obtained from Sigma Aldrich (St. Louis, MO, USA). Solutions of KML29, AM281, AM630, and Celecoxib were prepared in vehicle (1:1:18; DMSO:cremophor:saline) on the day of use. Rhodamine 6G (0.05%) and MIA were dissolved in saline. Physiological buffer (135 mM NaCl, 20 mM NaHCO₃, 5 mM KCl, 1 mM MgSO₄·7H₂O, pH = 7.4) was prepared in the lab.

Normal goat serum and lysophosphatidic acid (LPA) were obtained from Abcam (Toronto, ON, Canada), PF05089771 from Tocris Bioscience (Toronto, ON, Canada), AM281 from Cayman Chemical Company (Ann Arbor, MI, USA), PEP-1 from Abbiotec Inc. (San Diego, CA, USA), Fluoro-gold from Fluorochrome LLC (Denver, CO, USA), mouse IgG from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA), and guinea pig anti-Na_v1.7 (for IHC) from Alomone Labs (Jerusalem, Israel).
The mouse anti-Nav1.7 clone N68/6 monoclonal antibody used for behavioural experiments was developed by and obtained from the UC Davis/NIH NeuroMab Facility (Davis, CA, USA). The rabbit anti-guinea pig secondary biotin, streptavidin protein Dylight 488, and fluoromount G were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cremophor, dimethyl sulphoxide (DMSO), naloxone HCl and sodium monoiodoacetate (MIA) were obtained from Sigma-Aldrich (Oakville, ON, Canada). Isoflurane and euthansol were purchased from CDMV (Dartmouth, NS, Canada).