2.7. Determination of IL-1β. IL-1β was determined by using
AssayMax Human Interleukin-1β ELISA Kit (Catalog no.
E12200-1, St. Charles, MO 63304). A murine monoclonal
antibody specific for IL-1β has been precoated onto a
microplate. IL-1β in standards and samples was sandwiched
by the immobilized antibody and a biotinylated polyclonal
antibody specific for IL-1β, which was recognized by a
streptavidin-peroxidase conjugate. All unbound materials
were then rinsed off, and a peroxidase enzyme substrate
was added. The colour development was stopped, and
the intensity of the color was measured immediately at a
wavelength of 450 nm.