Mouse il 6 ready set go elisa kits

Mouse and Rat TNF-alpha Ready-SET-Go® ELISA kits, and Mouse IL-6 Ready-SET-Go® ELISA kits (eBioscience, USA) were used to assess the levels of TNFα and IL-6 in the macrophage activation experiments. Mouse CXCL1/KC DuoSet ELISA (R&D systems, USA) was used to measure mouse CXCL1 in all macrophage activation experiments. The procedures were carried out per the manufactures instructions, and all data were analyzed using the Epoch microplate spectrophotometer and Gen5 data analysis software (Biotek, USA).

A 20-gauge angiocatheter was ligated into the trachea, and the lungs were lavaged twice with sterile PBS (700 µL). The lavage fluid was centrifuged at 1,000 g for 10 min. The pellet was resuspended, and the cells were counted using the Invitrogen Countess Automated Cell Counter (Invitrogen, Grand Island, NY, USA). Protein levels in the supernatant were measured by Bradford Assay (BioRad), and cytokine levels were measured using ELISA. Interleukin (IL)-18 was measured using the mouse IL-18 ELISA Kit (MBL International Corporation, Woburn, MA, USA) according to the manufacturer’s instructions. Interleukin-6 (IL-6) was measured using the mouse IL-6 Ready-Set-GO ELISA Kits (eBioscience, San Diego, CA, USA). Interferon (IFN)-α was measured using the mouse IFN Alpha ELISA Kit (PBL Assay Science, Piscataway, NJ, USA).