Human elisa kit il 18 and il 21

The quantitation of cytokines was examined by BD OptEIA Set Human IFN-γ, TGF-β1, IL-10 (BD Bioscience) and Human ELISA Kit IL-18 and IL-21 (Invitrogen, USA). Briefly, target cells were cultured in a 6-well plate overnight and cocultured with effector cells (NK-92, ratio of E:T = 2:1) for 24 h with or without apigenin treatment (50 μM). After the coculture, the supernatants were harvested for ELISA assay. All experiments were conducted according to the manufacturer’s instructions. The absorbance of IFN-γ, TGF-β1 and IL-10 was measured at 450 nm with λ correction at 570 nm, and the absorbance of IL-18 and IL-21 was assessed at 450 nm with λ correction at 620 nm using a microplate reader (BMG Labtech, Germany).

The quantitation of cytokines was analyzed by BD OptiEIA Set Human IL-1β and TGF-β1 (BD Bioscience), and Human ELISA Kit IL-18 and IL-21 (Invitrogen, USA). Briefly, target cells were culutred at a density of 5 × 10⁵ cells per well of a 6-well plate overnight (18-24 h), and co-cultured with effector cells (NK-92) at a ratio of 2:1 (E:T) for 24 h with or without IL-6 antibody (2 ng/ml) or hrIL-21 (1 ng/ml). The supernatants co-cultured were harvested and stocked at -80°C before experiments. All experiments were performed in accordance with manufacturer’s instructions. The absorbances were measured at 450 nm with λ correction on 570 nm or 620 nm using a microplate reader (BMG Labtech, Germany).