Opteiatm set for mouse immunoglobulin e

Animals were euthanised via overdose with Sodium Pentobarbitone (200 mg/kg i.p.). Blood was attained via cardiac puncture with a heparinised syringe. Samples were then centrifuged (1398 g, 10 min, 4 °C) and the plasma supernatant transferred to eppendorfs and stored at −20 °C until subsequent analysis. The level of plasma IgE was used as a marker of allergic sensitisation and measured using Enzyme Linked Immunosorbant Assay (ELISA). Total and allergen specific IgE levels were measured using BD OptEIATM set for mouse immunoglobulin E (BD Biosciences, Oxford, UK) in accordance with the manufacturer’s instructions (see Additional file 1).

Isoflurane was from Abbott Laboratories (UK). BD OptEIATM set for mouse immunoglobulin E was from BD Biosciences (US). Medical Oxygen was from BOC Industrial Gases (UK). FBS and Fixable near-IR dead cell stain kit for 633/635 nm excitation was from Invitrogen (UK). Collagenase and DNase was from Roche Diagnostics (Germany). Alum^TM was from ThermoFisher Scientific (UK). Glycopyrrolate, ethanol and 2 N H₂SO₄ was from VWR (UK). Tiotropium was a gift from Boehringer Ingelheim (Germany). Sterile saline was purchased from Fresenius Kabi Limited (UK) and pentobarbitone from National Veterinary Services Limited (UK). All other agents were purchased from Sigma-Aldrich (UK) unless otherwise described.