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The PVDF membrane was subsequently washed (5 min 5) with tris-buffered saline with tween (TBST) buffer and then incubated with the appropriate secondary antibody (Peroxidase-conjugated goat anti-rabbit IgG (H + L), at a dilution of 1:5000) in a 5% bovine serum albumin solution for 1 h at 25 C

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Intensity ratios between the paramagnetic and diamagnetic states were computed for all assigned and non-overlapping 1 H, 15 N, and 1 H, 13 C HSQC cross-peaks, employing a correlation time of the electron-spin interaction vector of 4 ns 52,53

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