Human il 6 quantikine enzyme linked immunosorbent assay elisa kit

The cells (2 × 10⁵) were seeded in a 60 mm tissue culture dish (Asahi Techno Glass Co., LTD, Shizuoka, Japan) and incubated in DMEM with 1% FBS for 24 h. The cells were then irradiated with 6 and 10 Gy of X-rays. At 12, 24, 36, 48, and 60 h after irradiation, the conditional culture media were collected, and the concentration of IL-6 was measured using a Human IL-6 Quantikine enzyme-linked immunosorbent assay (ELISA) Kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer's instructions.

IL-6 levels were measured using a Human IL-6 Quantikine enzyme-linked immunosorbent assay (ELISA) kit (Bio-Techne-R&D Systems Inc., Minneapolis, MN, USA) employing a sandwich-type immunoassay, as previously described.7 (link) After pipetting the reference standard and samples into the wells of the microplate, which was precoated with monoclonal antibodies for human IL-6, the immobilized antibody bound to any IL-6 present. Any unbound substances were washed away, and enzyme-linked polyclonal antibodies were added. After washing, a substrate solution was added to detect the amount of IL-6 bound. After adding the stop solution, the color intensity was measured, and the absorbance was examined at 450 nm using a Spectra Max 340 pc (Molecular Devices Co., CA, USA). The results were reported as the amount of IL-6 protein in 1 mL supernatant.