The use of fluorescence-based detection of single biomolecules under physiological conditions has opened new doors for studying biological problems. Fluorophore bioconjugation technologies enable target visualization and rapid probing of proteins, and thus have become a major tool in biological sciences. Most commonly, amine-reactive fluorophores such as NHS esters are used to react with free amines present at either N-terminus or lysine side chains of the target protein, resulting in a stable covalent linkage. Recent development in bioconjugation chemistry has made it possible to target amino acid sides other than lysine.

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