The ProQuantum Human IL-12p40 Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-12p40 protein in small sample volumes. Utilizing proximity ligation assay (PLA) technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive Cloud-based software for analytics enables sample-to-answer in just 2 hours.
• High sensitivity—detect low levels of protein with greater sensitivity than traditional methods
• Broad dynamic range—≥5 logarithmic units, minimizing sample dilutions to ensure they fall within the range
• Small sample consumption—use 2–5 µL of sample (compared to 75 µL for triplicate wells with other methods)
• Fast, easy workflow—2 hours from sample to answer, with no wash steps
• No proprietary instrument to purchase—runs on any real-time PCR instrument
ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology (Figure 1).
The assay workflow is fast and easy—2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum cloud-based software at https://apps.thermofisher.com/apps/proquantum. Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, set up customized assay instructions, and obtain robust statistical group-wise comparisons.
Interleukin-12 (IL-12) is a heterodimeric 70 kD (p70) cytokine composed of two covalently linked, glycosylated chains, 40kD (p40) and 35-kD (p35). IL-12 is mainly produced by monocytes, macrophages, and dendritic cells in response to bacterial products such as lipopolysaccharides (LPS), to intracellular pathogens or upon interaction with activated T cells. IL-12 was originally discovered because of its ability to induce interferon-gamma production, cell proliferation, and cytotoxicity mediated by natural killer cells and T cells. It is now established that IL-12 also plays a key role in the development of Th1 responses, leading to IFN-gamma and IL-2 production. These cytokines can in turn promote T-cell responses and macrophage activation. The p40 and p35 subunits by themselves have no IL-12 bioactivity, though the p40 homodimer has been shown to bind the IL-12 receptor and to be an antagonist of IL-12 p70. Free p40 is typically secreted in vast excess of IL-12 p70 by cells co-expressing both the p35 and p40 subunits. The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the IL-6 receptor, while the p35 subunit shows some homology to IL-6 and G-CSF.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.