Tnf α and il 1β elisa kits

TNF-α and IL-1β ELISA kits were purchased from Biolegend (CA, USA). Tissue protein extract and BCA Protein Assay Kit were bought from Thermo (Thermo, MA, USA). Trypticase Soy Broth (TSB) and bacillus medium were purchased from Qingdao Haibo Biotechnology Co., Ltd (Qingdao, China). Crystal Violet Stain solution, 1% was acquired from Solarbio (Solarbio, Beijing, China). All the monoclonal antibodies, including β-actin, p65, IκB, the phosphorylation of p65, phosphorylation IκB, ZO-1, Occludin, and Claudin-3 were recruited from Cell Signaling Technology (Beverly, MA, USA). MPO kit was available from Nanjing Jiancheng Co., Ltd (Nanjing, China). Animals A total of 60 Balb/c mice (40 females and 20 males) aged 6-8 weeks were purchased from the Liaoning Changsheng Biotechnology Co. Ltd. The animal experiments were subject to approval by the Animal Ethics Committee of Jilin University (KT202103058). Females and males are mixed in miniature isolation cages in about a ratio of 2 to 1 after adapting to the environment with free food and water. The mastitis model was established complying with the experimental animal manual. This study is built on the Handbook on the Care and use of Experimental Animals published by the National Institutes of Health.