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Efforts to elucidate the roles of protein sulfenylation in physiology and disease have been hampered by the lack of techniques to probe these modifications in native environments with molecular specificity. In this review, we trace the history of chemical and biological methods that have been developed to detect protein sulfenylation and illustrate how a recent cell\u2010permeable chemical reporter, DYn\u20102, has been used to detect and identify intracellular targets of endogenous H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> during growth factor signaling, including the epidermal growth factor receptor. The array of new tools and methods discussed herein enables the discovery of new biological roles for cysteine sulfenylation in human health and disease. \u00a9 2013 Wiley Periodicals, Inc. 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