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Detection of in-cell protein sulfenylation with genetically encoded Yap1-cCRD probe. Cells expressing Yap1-cCRD are exposed to peroxide and protein conjugates are extracted with trichloroacetic acid (TCEP). Free thiols are capped by alkylation with iodoacetamide (IAA). Yap1-cCRD-captured proteins are affinity enriched and eluted with reducing agents (DTT or TCEP). After sample enrichment and protease digestion, the resulting peptides are analyzed and identified by LC–MS/MS.
