Supplementary Figure 5: The effect of crowding induced by bovine serum albumin (BSA) on the sensor readout. | Nature Methods

Supplementary Figure 5: The effect of crowding induced by bovine serum albumin (BSA) on the sensor readout.

From: A sensor for quantification of macromolecular crowding in living cells

Supplementary Figure 5

(a) Emission spectrum upon excitation of mCerulean at 420 nm at different % w/w BSA. The relative contribution of mCitrine increases according to an increase in crowding, but the overall fluorescence decreases. The decrease is most likely due to an inner filter effect, because the BSA solution is strongly absorbing at 420 nm. Conditions: 10 mM NaPi, 100 mM NaCl, pH 7.4. (b) Emission spectrum upon excitation of mCitrine at 515 nm at different % w/w BSA. An approximately linear decrease in intensity is observed upon increasing the concentration of BSA. The decrease is likely due to the inner filter effect, because the BSA solution absorbs at 515 nm (most likely due to traces of myoglobin). Conditions: 10 mM NaPi, 100 mM NaCl, pH 7.4. (c) The FRET ratio, calculated from Supplementary Figure 5a, plotted versus the concentration of BSA. The FRET ratio increases with increasing concentration of BSA. The error bars are standard deviations from three independent experiments.

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