N-hydroxysuccinimide (NHS) prolongs the lifetime of amino-reactive moites formed at carboxylic acids and increases the efficiency of the zero-length crosslinker EDC. During NHS crosslinking, NHS at concentrtion of 5 mM reacts with EDC to yield a semi-stable NHS ester, that subsequently can be reacted with primary amines (-NH2), resulting in amide crosslinks. NHS is soluble in aqueous and organic solvents, however NHS activation may decrease water solubility of the modified carboxylate molecule. Sulfo-NHS can preserve and even increase the water solubility of targeted molecules, due to the presence of the charged sulfonate moity.

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