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After tris-buffered saline with tween 20 (TBST) washes, membranes were incubated with corresponding secondary antibody conjugated with horseradish peroxidase (1:5000 dilution in TBST) for 1 h at room temperature

While HDX-MS is characterized by somewhat lower spatial resolution than HDX-NMR [although there are several computational strategies to extract single-residue protection factors from peptide-level HDX-MS data (Zhang et al., 2012