Description
Mouse IL-12p70 is a heterodimeric pro-inflammatory cytokine composed of the p35 and p40 subunits, and belongs to the IL-12 cytokine family, which also includes IL-23, IL-27, and IL-35. The p35 subunit also associates with EBI3 to form IL-35, whereas the p40 subunit can form IL-23 (in combination with p19) or exist as a p40 homodimer (IL-12p80) or monomer. These different combinations confer distinct and sometimes opposing biological functions within the IL-12 family.
IL-12p70 is primarily produced by antigen-presenting cells such as dendritic cells, macrophages, and neutrophils, and can also be secreted by activated B cells and helper T cells in response to antigenic stimulation and inflammatory signals. Functionally, IL-12 plays a central role in bridging innate and adaptive immunity. It promotes the differentiation of naive CD4⁺ T cells into Th1 cells, enhances interferon-γ (IFN-γ) production, and stimulates the cytotoxic activity of natural killer (NK) cells and CD8⁺ T lymphocytes.
These effects are mediated through binding to the heterodimeric IL-12 receptor (IL-12Rβ1 and IL-12Rβ2), leading to activation of the JAK/STAT signaling pathway, particularly STAT4.
Studying the production and regulation of IL-12p70 during infection is therefore critical for understanding immune defense mechanisms and the orchestration of inflammatory responses.
Formats
Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Features
No wash steps, no separation steps
Alternative to ELISA technology
Sensitive detection
Broad sample compatibility
Small sample volume
Results in less than 4 hours
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules, triggering a cascade of energy transfer in the Acceptor beads, and resulting in a sharp peak of light emission at 615 nm.
Principle of the AlphaLISA Mouse IL12p70 assay
The AlphaLISA Mouse IL12p70 assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-mIL12p70 antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-mIL12p70 antibody conjugated to AlphaLISA Acceptor beads. One antibody is directed against the mouse p35 subunit protein, and the second recognizes specifically the p40 subunit forming mIL12p70. In the presence of the target, both antibodies bind to mouse IL12p70, and the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules, triggering a cascade of energy transfer within the Acceptor beads, and resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of mIL12p70 present in the sample (cell supernatant or plasma EDTA).
Protocol of the AlphaLISA Mouse IL12p70 assay
The AlphaLISA Mouse IL12p70 assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples (cell supernatants or plasma EDTA) or standards are dispensed directly into the assay plate for the detection of Mouse IL12p70 by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
