1. Peptide and Protein Synthesis:

As a carboxyl activator and dehydrating agent, it efficiently promotes the formation of amide bonds between amino acids in peptide synthesis, commonly used in solid/liquid phase peptide chain assembly; it is also used for protein fragment linkage and modification, such as introducing special functional groups. It is often used in combination with HOBt, Sulfo-NHS, etc., to inhibit racemization and improve reaction efficiency.

2. Biomolecular Crosslinking and Labeling:

Used for zero-length crosslinking of proteins-proteins, proteins-nucleic acids, and proteins-carriers (such as BSA, KLH), for studying molecular interactions, preparing immunogens, or stabilizing enzyme-substrate complexes; it can also achieve covalent coupling of antibodies with fluorescent dyes, biotin, and other labels for immunoassay, imaging, and diagnostic reagent preparation.

3. Nucleic Acid Chemistry:

It can react with the 5′ phosphate group of nucleic acids, and after activation, it is used for coupling nucleic acids with proteins, small molecules, or solid-phase carriers, such as preparing nucleic acid probes, immobilizing DNA/RNA chips, and linking carriers to nucleic acids in gene delivery systems.

4. Biopharmaceutical R&D:

Used for the synthesis and modification of peptide drugs, antibody-drug conjugates (ADCs), and peptide vaccines, constructing stable amide bonds to ensure bioactivity; also used for the coupling of polymers and target molecules in drug delivery systems to achieve targeted drug delivery.

5. Material Surface Functionalization:

As a coupling agent between carboxyl and amino groups, it immobilizes biomolecules (such as enzymes and antibodies) on the surfaces of polymers, glasses, and nanoparticles to prepare biosensors, bioactive coatings for medical implants, diagnostic chips, etc., endowing materials with biorecognition or catalytic functions.

6. Organic Synthesis Applications:

Used in the preparation of amides, esters, ureas, and other compounds in small molecule organic synthesis, such as the synthesis of pharmaceutical intermediates; it can also catalyze the esterification of carboxylic acids with alcohols and the amidation of carboxylic acids with amines, suitable for water-soluble or water-organic mixed systems, with mild reaction conditions (pH 4.0-6.0, amine-free/carboxylate buffer).

7. Biochemical Detection and Analysis:

In experiments such as enzyme-linked immunosorbent assay (ELISA) and Western blotting, it is used for antigen-antibody cross-linking, signal molecule coupling, or activation and coating of solid-phase carriers to improve detection sensitivity and specificity.