This chemical is widely used as a chromogenic substrate in biochemical assays, particularly in the detection of β-glucuronidase enzyme activity. It is commonly employed in microbiology, molecular biology, and histochemistry to identify and study bacterial colonies, such as E. coli, which express β-glucuronidase. Upon enzymatic cleavage, it produces an insoluble, colored indigo derivative, allowing for easy visualization of enzyme activity. This makes it a valuable tool in diagnostic testing, environmental monitoring, and research applications where precise detection of enzyme presence is critical.