The ProQuantum Human IL-6 Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-6 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 6 (IL-6) is a multifunctional 26 kD protein originally discovered in the medium of RNA-stimulated fibroblastoid cells. IL-6 appears to be directly involved in the responses that occur after infection and cellular injury, and it may prove to be important as IL-1 and TNF-a in regulating the acute phase response. IL-6 is reported to be produced by fibroblasts, activated T cells, activated monocytes or macrophages and endothelial cells. IL-6 acts upon a variety of cells including myeloid progenitor cells, T cells, B cells, and hepatocytes. In addition, IL-6 appears to interact with IL-2 in the proliferation of T lymphocytes. IL-6 potentiates the proliferative effect of IL-3 on multipotential hematopoietic progenitors. IL-6 plays a critical role in B-cell differentiation to plasma cells and is a potent growth factor for plasmacytoma and myeloma. IL-6 is a very useful culture supplement for the generation of a high number of antibody-producing hybridomas. Primarily produced at sites of acute and chronic inflammation, IL-6 is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of IL-6 is implicated in a wide variety of inflammation-associated disease states including diabetes mellitus and systemic juvenile rheumatoid arthritis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.