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The labeled protein concentrations were adjusted to 100 nM based on fluorescence intensity measured during pretest scans, ensuring signal quality and uniformity across all three Syn constructs for direct comparison of binding affinities

Post-translational modifications observed in Prx2, such as phosphorylation, proteolysis, and even the hyperoxidation of its peroxidatic cysteine, are involved in the regulation of its peroxidase activity and could affect its participation in redox signaling pathways [87,88,89]

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In one example, the T7 RNA polymerase-promoter system is used and is induced from the lac operon in BL21 (DE3)-based strains to circumvent the need to separately purify and supplement T7 RNA polymerase into CFE reactions