Description
Principle of the assay: human IL-12(P70) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-12(P70) has been precoated onto 96-well plates. Standards(sf21,(I23-S328)+(R23-S219)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-12(P70) is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human IL-12(P70) amount of sample captured in plate.
Background: Interleukin (IL)-12 is a 70-KDa cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response.i And the IL-12p70 is a heterodimer of p35 and p40 subunits; it is animportant cytokine secreted by antigen-presenting cells in response to antigenic stimulation.ii Gene expressionanalysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40 (protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT- 18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70.iii The standard product used in this kit isrecombinant human IL-12 p70 with the molecular mass of 75 KDa