5-ROX is widely used as a fluorescent dye in molecular biology and biochemistry. It is particularly valuable in real-time PCR (polymerase chain reaction) applications, where it serves as a reporter dye for detecting and quantifying nucleic acids. Its fluorescence properties make it suitable for use in multiplex assays, allowing simultaneous detection of multiple targets. Additionally, 5-ROX is employed in fluorescence microscopy and flow cytometry for cellular imaging and analysis. Its stability and brightness enhance the accuracy and sensitivity of these techniques, making it a preferred choice in research and diagnostic applications.