Mouse rat il 33 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 the quantification of sST2, the cells were cultured for 24 h, and the conditioned media were collected. The amount of sST2L was measured using a mouse ST2L/IL-1 R4 Quantikine ELISA kit (R&D Systems) according to the manufacturer's instructions. For the quantification of IL-33 in tumour tissues, dissected tissues were homogenised in cold DPBS with a Potter-Elvehjem homogeniser, sonicated and cleared by ultracentrifugation at 45 000xg for 30 min. The resulting supernatant was used to quantify the amount of IL-33/g of tumour tissue wet weight using a mouse/rat IL-33 Quantikine ELISA kit (R&D Systems). For the quantification of IL-33 secreted by IL-1β-treated P29 cells, the cells (1 × 10⁶ cells) were cultured for 3 days, and the conditioned media were collected and concentrated 10-fold using Amicon Ultracel-3 K (Millipore, Billerica, MA, USA). IL-1β-treated P29 cells were lysed in RIPA buffer and cleared by centrifugation at 10 000xg for 20 min. The resulting supernatant was used to quantify the amount of IL-33 and total protein.