Mouse il 15 il15r platinum elisa

For IL-15, the ELISA assay used to measure the concentration of bioactive IL-15 and IL-15:IL-15Rα in culture supernatants. In shorts, 1 × 10⁶ cells were incubated in 1 ml of culture medium in 24-well culture plates at 37°C for 24 h. IL-15 or IL-15:IL-15Rα complexes in the culture supernatants was measured with Mouse IL-15 DuoSet ELISA kit (DY447-05; R&D System, USA) or mouse IL-15/IL15R Platinum ELISA (BMS6023; Affymetrix, USA) respectively. For interferon γ (IFN-γ) assay, control and surviving mice from vaccination experiment were injected with 1 × 10⁶ MMC inactivated wild-type CT26 cells and splenocytes were collected after 1 week. Each suspension of splenocytes (2 × 10⁶ cells/well) was seeded with 600 μl complete culture media (RPMI; 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, 50 μM/ml of 2-ME) and treated with cancer cell lysate (2 × 10⁵ boiled CT26 cells/well). After 72 h, 100 μl of culture supernatant from those stimulated splenocytes were subjected to IFN-γ ELISA (IFN-γ ELISA Kit; BioLegend, USA).