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N-Hydroxysuccinimide (NHS) is an organic compound with the molecular formula C4H5NO3C4​H5​NO3​ and a CAS number of 6066-82-6. It appears as a white solid and is primarily used as a reagent in organic chemistry, particularly for preparing active esters in peptide synthesis and bioconjugation techniques. Chemical Properties Molecular Weight: 115.09 g/mol Melting Point: 93 - 100 °C4. Solubility: Soluble in water2. Synthesis NHS can be synthesized by heating succinic anhydride with hydroxylamine or hydroxylamine hydrochloride. This process yields NHS, which can then be utilized to activate carboxylic acids, facilitating their reaction with amines to form amides, a crucial step in peptide synthesis5. Applications NHS is widely employed in various applications: Peptide Synthesis: It is used to create NHS esters from carboxylic acids, which are then used to couple with amines to form peptides3. Protein Modification: NHS esters are commonly used for labeling proteins with fluorescent dyes or enzymes, enhancing their utility in biochemical assays5. Biosensor Development: NHS can be combined with coupling reagents like EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) for enzyme immobilization in biosensors3. Alternatives There are several alternatives to NHS for similar applications: Sulfo-NHS: A water-soluble analog. Hydroxybenzotriazole (HOBt) and 1-Hydroxy-7-azabenzotriazole (HOAt): Other reagents used for activating carboxylic acids2