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. 2018 Oct 18;6:146. doi: 10.3389/fbioe.2018.00146

Figure 2.

Figure 2

Fluorescence response of the MTase assay. To assess the response of the MTase assay, a 100 μl reaction typically contained the following assay components in 50 mM potassium phosphate buffer (pH 7.5): 3.5 μg purified recombinant Mtn, 1 μg XOD (10 units/mg), 0.2 μg HRP (300 units/mg), and 100 μM of the substrate. Fluorescence response was monitored in 96-well microplates with excitation and emission wavelengths set to 530 and 590 nm, respectively, in the presence of (A) 100 μM hydrogen peroxide, 100 μM adenine, 100 μM SAH and in the absence of the aforementioned chemicals, or (B) the presence and absence of the assay-coupling enzymes.