The ProQuantum Human IL-17F Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-17F 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 below).
The assay workflow is fast and easy—2 steps in 2 hours. There are a total of 7 components in each kit (figure below). 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.
IL-17F (Interleukin 17F, CTLA-8)) is a cytokine belonging to the IL-17 family that is produced by inflammatory cells such as activated T cells, mast cells, and basophils. IL-17F is involved in allergic airway inflammation, and can induce several cytokines, chemokines, and adhesion molecules in bronchial epithelial cells, vein endothelial cells, fibroblasts, and eosinophils. IL-17F may be secreted as a homodimer, or a heterodimer with IL17A. It acts by binding to the type I receptor, IL-17R, aiding recruitment of monocytes and neutrophils at the site of inflammation by increasing chemokine production. IL-17F also stimulates induction of other pro-inflammatory cytokines TNF alpha, IL-1 beta, IL-6, and IL-8, and reports strongly suggest the involvement of IL-17 in several chronic inflammatory diseases such as rheumatoid arthritis, psoriasis and multiple sclerosis. TGF-beta (differentiation) and IL-23 (expansion) are required for induction and maintenance of Th17 (IL-17 producing) cells, which in turn induce the other pro-inflammatory cytokines. IL-17F is produced, and exists, as a homo-dimer, with homology to a herpes virus early protein, is one of the six members (IL-17A-F) of this cytokine family, and is well characterized and highly expressed by activated effector memory T cells. IL-17F has been found to inhibit the angiogenesis of endothelial cells and induce endothelial cells to produce IL2, TGFB1/TGFB, and monocyte chemoattractant protein-1.
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