Hplc grade

BALB/c mice (specific pathogen-free grade, aged 5-6 weeks, weight 16-20 g) were purchased from the Animal Center of Guangdong Province (Guangzhou, China) and kept in a specific pathogen-free grade breeding room. This study was carried out strictly in accordance with the recommendations of the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Committee on the Ethics of Animal Experiments of the Shenzhen University. Silkworm pupae were donated by the Sericulture and Agro-Processing Research Institute, Guangdong Academy of Agricultural Sciences (Guangzhou, China). Reagents and chemicals were analytical grade (Sigma, St Louis, MO, USA) and solvents were HPLC grade (Mallinckrodt, Lane Cove West, NSW, Australia). SDS/ PAGE/western blotting was performed by using a Mini Protean II apparatus (BioRad, Hercules, CA, USA) with 12 or 15% gels and a Tris/Tricine buffer system. Two dimensional PAGE was performed by using Immobiline IPG strips (13 cm) with a pH range of 3-10 (GE Healthcare Bio-Science, Piscataway, NJ, USA). Methacholine was purchased from Sigma-Aldrich (St Louis, MO, USA) and biotin-labeled goat anti-mouse IgE from Zymed Laboratories (San Francisco, CA, USA).