Widely used in molecular biology and histochemistry as a chromogenic substrate for alkaline phosphatase detection. When cleaved by alkaline phosphatase, it produces a blue-purple insoluble precipitate at the site of enzyme activity, enabling visualization of gene expression or protein localization in techniques such as in situ hybridization, immunohistochemistry, and blotting assays. Its combination with other substrates allows for multiplex staining, where different enzymes or targets can be distinguished by color. The presence of bromo and chloro substituents enhances the stability and specificity of the color development. Commonly used in developmental biology and pathology research for precise spatial and temporal detection of enzymatic activity.