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Reduction/Alkylation Tradeoffs Denaturant-Induced Modifications: Optimized Protocol: Reduction: 5mM TCEP (30min, 25C) Alkylation: 10mM CAM (20min, 4C darkness) Minimizes side reactions 6

Nat Biotechnol 36: 880887 Colaert N, Helsens K, Martens L, Vandekerckhove J, Gevaert K (2009) Improved visualization of protein consensus sequences by iceLogo