Mouse st2 il 1 r4 quantikine elisa kit

Conditioned media were harvested after 24 h of culture and centrifuged at 15,000 × g for 20 min at 4°C. For serum preparation, blood was allowed to clot and centrifuged for 10 min at 1,000 g. sST2 and IL-33 concentrations were measured with the Mouse ST2/IL-1 R4 Quantikine ELISA Kit (R&D Systems) and the Mouse/Rat IL-33 Quantikine ELISA Kit (R&D Systems), respectively, according to the manufacturer’s instructions.

For serum preparation, blood was allowed to clot and then centrifuged for 10 min at 1,000g. Serum was collected and immediately stored at −80 °C prior to use. For the preparation of conditioned media, the cells were seeded at a concentration of 1 × 10⁶ cells per well in a 24-well plate (BD Falcon, Franklin Lakes, NJ, USA) in 0.5 ml of Dulbecco's modified Eagle's medium containing 1% FBS. Conditioned media were harvested after 24 h of cultivation. For the preparation of tissue lysates, tissues were cut into pieces using a razor blade and were then placed in ice-cold DPBS containing cOmplete Protease Inhibitor Cocktail (Roche) and homogenized for 3 min at 30 Hz using a TissueLyser (QIAGEN). Supernatants were collected after centrifugation at 15,000g for 20 min at 4 °C. Human and mouse sST2 concentrations were measured with the Human sST2 (soluble ST2) ELISA Kit (MBL) and the Mouse ST2/IL-1 R4 Quantikine ELISA Kit (R&D Systems), respectively, according to the manufacturer's instructions. Mouse VEGF concentrations were measured with the Mouse VEGF Quantikine ELISA Kit (MBL). The concentrations of interferon-γ (IFN-γ), IL-4, IL-13 and IL-33 were measured with Ready-Set-Go ELISA kits (e-Bioscience, San Diego, CA, USA).