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A Quantum Leap in mRNA Quantitation

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

By | October 30, 2000

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 using an anti-digoxigenin alkaline phosphatase conjugate and NADPH substrate solution.

The Quantikine® mRNA assay is less labor-intensive than most other methods of RNA quantitation yet provides the sensitivity of a northern blot and does not require the use of a radioisotope as an RPA often does. In addition, the microplate format permits the user to analyze up to three different gene-specific targets at once, all in less than five hours.

Product manager Leena Martel says that she has received consistently good reviews from customers using the assay and explains that "the simplicity of the Quantikine mRNA assay is the most attractive feature of this kit."

Nasser Chegini, professor of reproductive endocrinology at the University of Florida, has found the Quantikine mRNA assay to be "rapid, accurate, and less labor-intensive than competitive quantitative RT-PCR." Currently, there are about 20 gene-specific mRNA kits available (human and mouse cytokines).

--Hillary E. Sussman (hes01@health.state.ny.us)

For More Information
R&D Systems, Inc
(800) 343-7475
www.rndsystems.com

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