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. 2014 Oct 1;12(8):481–496. doi: 10.1089/adt.2014.603

Fig. 5.

Fig. 5.

Triton X-100 tolerance. (A) Triton X-100 (0.01% v/v) has no discernible effect upon fluorescence intensity measurements of 3-O-methylfluorescein (OMF) when path length is fixed by using a cuvette. Fluorescence emission scans (485-nm excitation) were performed in quartz cuvettes on solutions of 1 μM OMF prepared in reaction buffer with (▼) and without (▲) the presence of 0.01% Triton X-100. Emission spectra for each solution are plotted together on the same graph and are almost perfectly superimposable. (B) Triton X-100 (0.01% v/v) has no significant effect upon fluorescence intensity measurements for PP5C reaction progress curves when path length is fixed. PP5C (1 nM) was incubated as described in “Materials and Methods” section with 50 μM OMFP in assay buffer with (▼) and without (▲) the presence of 0.01% Triton X-100. Fluorescence intensity measurements of the reaction mixture in a quartz cuvette were conducted every 3 s for 5 min (485-nm excitation/525-nm emission) with the M5. Reaction progress curves were plotted together (adjusting starting fluorescence intensity to zero) with GraphPad Prism and are almost perfectly superimposable. (C) Triton X-100 (0.01% v/v) has no significant effect upon fluorescence intensity measurements for PP1α reaction progress curves when path length is fixed. PP1α (6.67 nM) was incubated as described in “Materials and Methods” section with 100 μM OMFP in assay buffer with (▼) and without (▲) the presence of 0.01% Triton X-100. Fluorescence intensity measurements of the reaction mixture in a quartz cuvette were conducted every 3 s for 5 min (485-nm excitation/525-nm emission) with the M5. Reaction progress curves are plotted together (adjusting starting fluorescence intensity to zero) with GraphPad Prism and are almost perfectly superimposable.