The NWLSS™ Human IL-12p40 (Subunit β) ELISA kit is intended to be used for the in vitro quantitative determination of the p40 or β subunit of human interleukin 12 in human serum, plasma, cell lysates and cell culture supernatants. The assay will recognize native and recombinant human IL-12p40.
Interleukin-12 (IL-12p70) is a cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response. The IL-12p70 heterodimer is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation. Functional IL-12p70 acts as a pro-inflammatory cytokine to stimulate T-cells and natural killer (NK) cells to produce gamma interfeuron (IFN-gamma). However, IL-12 p40 (subunit β) has been shown to also play an independent regulatory role by competing for binding with the IL-12 receptor and also an independent pro-inflammatory role as a chemoattractant for macrophages and stimulated dendritic cells. It is now known that IL-12p40 can be secreted as either a monomer (IL-12p40) or as a homodimer (IL12p80) which seems to function more as a pro-inflammatory cytokine. The standard product used in this kit is recombinant human IL-12p40 subunit with the molecular mass of 40 KDa.
The NWLSS™ Human IL-12p40 ELISA is a sandwich format Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to human IL-12p40. Samples are pipetted into these wells. Non-bound IL-12p40 and other components of the sample are removed by washing, after which a biotinylated antibody specific to human IL-12p40 is added. In order to quantitatively determine the amount of IL-12p40 present in the sample, Streptavidin Horseradish Peroxidase (HRP) conjugate is added to each microplate well. After another wash step, TMB-substrate solution is added to each well. Finally, a sulfuric acid stop solution is added and the resulting yellow colored product is measured at 450nm. The amount of sample IL-12p40 is determined by direct comparison with the standard curve generated in the assay.
View Assay Protocol
1 X 96 well ELISA presented as 12 X 8 well (6 X 16 well) strips in frame.
For quantitative detection of human IL-12p40 (IL-12 Beta Subunit)
Human serum, plasma, cell lysates and cell culture supernatants.
|Storage and Stability:||
9 months from date of manufacture
1 Foil Pouch 96 well microplate precoated with anti-Human IL-12p40|
2 bottles 20X Concentrated Wash Buffer (25 mL)
1 vial rHu IL-12p40 Standard (lyophilized) (1 Vial)
1 bottle Standard/Sample Dilution Buffer (25mL)
1 vial Secondary Antibody (Lyophilized) (1 Vial) (Biotinylated Anti-Hu IL-12p40)
1 vial 100X Streptavidin-HRP Conjugate (150 µL)
1 bottle Reagent Dilution Buffer (25mL)
1 bottle TMB Substrate (15 mL)
1 bottle Stop Solution (1 N Sulfuric Acid, H2SO4) (15 mL)
2 Adhesive Plate Covers (2)