2-Nitro-5-thiocyanatobenzoic acid (NTCB) is a versatile reagent primarily utilized in organic synthesis for introducing thiocyanate functional groups into various compounds. It is particularly valuable in the preparation of heterocyclic compounds, where the thiocyanate group serves as a precursor for further chemical transformations. Additionally, it finds application in the development of pharmaceuticals and agrochemicals, contributing to the synthesis of biologically active molecules. Its nitro group makes it a useful intermediate in the production of dyes and pigments, enabling the creation of complex aromatic systems.

In biochemical research, NTCB is widely used for the specific cleavage of peptide bonds on the N-terminal side of cysteine residues in proteins, facilitating structural analysis and modification of biomolecules. This property, stemming from its reactive nitro and thiocyanate groups, makes it essential in studies involving protein-protein interactions, enzyme assays, and proteomics.