Cytokines, unique growth factors that are secreted by various cells of the body, include interleukins (ILs), interferons (IFNs), and tumor necrosis factors (TNFs). These proteins bind to target cell surface receptors and activate cell proliferation and/or differentiation. Until recently, research that quantifies levels of cytokine-specific mRNA using traditional methods such as the northern blot, slot blot, quantitative RT-PCR, and ribonuclease protection assay (RPA) has been labor-intensive and time-consuming. Additionally, these methods are either unsuitable for high-throughput analyses or require the use of a radioisotope.
R&D Systems' Quantikine mRNA kit R&D Systems Inc., of Minneapolis, has developed a novel method for quantitation of cytokine-specific mRNA using a colorimetric microplate assay. Total or poly-(A)+ RNA samples are first hybridized with gene-specific biotin-labeled oligonucleotide probes and digoxigenin-labeled detection probes. The RNA/probe hybrid then is captured on transfer to a streptavidin-coated microplate, and color is developed in proportion to the amount of gene-specific mRNA ...
