RNA plays crucial role in coding, decoding and regulation of genes, and protein expression in all living cells. The ability to detect newly synthesised RNA or changes in RNA levels under various physiological conditions, or resulting from disease, environmental damage, or drug treatments is an important aspect of toxicological profiling. Many anti-cancer drugs inhibit transcription, and most transcription inhibitors have useful pharmacological properties. BioVision’s EZClick™ Global RNA Synthesis Assay Kit provides a simple and robust tool for detection of global RNA transcription, temporally and spatially, or changes in RNA levels directly in living cells. De novo synthesised RNA can be detected with a simple procedure without the use of radiolabeling or antibodies. The kit principle relies on the incorporation of cell permeable 5-EU (Ethynyl uridine) into nascent RNA, but not into DNA, instead of its natural uridine analog. 5-EU can be used as a replacement for BrU (5-Bromo-uridine) to measure de novo synthesised RNA in proliferating cells. Modified RNA is detected by click chemistry using an azide-containing dye that enables for multiplex analyses with other probes, or detection of RNA-interactive proteins for deeper biological insights. The kit provides sufficient materials for 100 assays for quantitative analysis by microplate reader (Ex/Em= 494/521 nm) and/or by fluorescence microscope. The kit includes Actinomycin D, an inhibitor RNA synthesis that serves as an experimental control.
Biochemical and cellular assays