Application note: High throughput quantitation of cytokine biomarkers using LANCE Ultra TR-FRET assays
In this application note, PerkinElmer demonstrates the sensitivity, robust performance, and HTS suitability of the LANCE Ultra cytokine biomarker kits.
Tissue injury can lead to inflammation, which is characterised by an accumulation of cytokines. The excessive production of pro-inflammatory cytokines, such as TNFα, IL-6 and IL-1β, can result in further tissue damage and has been implicated in many diseases, such as atherosclerosis and cancer. Anti-inflammatory drug development is therefore an important area of focus in the pharmaceutical industry.
Discovery of these drugs through high throughput screening campaigns requires the ability to both robustly and reliably detect cytokine production in a variety of complex sample matrices.
LANCE® and LANCE (Lanthanide chelate excite) Ultra are our TR-FRET (time-resolved fluorescence resonance energy transfer), homogeneous (no wash) technologies. One protein of interest is labelled with a donor fluorophore (a LANCE Europium chelate) and the second protein is labelled with an acceptor fluorophore (ULight™ dye). Upon excitation at 320 or 340 nm, energy can be transferred from the donor Europium chelate to the acceptor fluorophore if sufficiently close for FRET (~10 nm). This results in the emission of light at 665 nm
Here we demonstrate the ease and versatility of using LANCE Ultra TR-FRET assays to quantitate low levels of the pro-inflammatory cytokines TNFα, IL-6, and IL-1β and to detect measurable inhibition of cytokine secretion using the small molecule dexamethasone. Furthermore, we show the accuracy of miniaturisation of the assay, demonstrating how it is suitable for high throughput screening.
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