The Human IL-33 Cytokine domain Detection Kit measures human IL-33 by sandwich ELISA. The assay uses polyclonal antibody and monoclonal antibody against two different epitopes of human IL-33.
In the wells coated with anti-human IL-33 polyclonal antibody, samples to be measured or standards are incubated. After washing, the biotinylated anti-human IL-33 monoclonal antibody is added into the microwells and incubated. After another washing, the streptavidin-HRP is added and incubated. Following a final washing, the peroxidase substrate is mixed with the chromogen and allowed to incubate for an additional period of time.
An acid solution is then added to each well to terminate the enzyme reaction and to stabilize the developed color. The optical density (O.D.) of each well is then measured at 450 nm using a microplate reader. The concentration of human IL-33 is calibrated from a dose response curve based on reference standards.