Analytica Chimica Acta, accepted, 07/07/2015. This is the accepted version without proofing
corrections. DOI: 10.1016/j.aca.2015.06.011
.
Page 8 of 26
Figure 3: MCR scores plots and normalized MCR scores plots of the pure components extracted from HH (a/b),
VV (c/d), VH (e/f), and HV (g/h) polarized TSFS spectra of 1 mg mL
–1 HSA in PBS undergoing thermal
unfolding from 1070 °C and cooling from 7020 °C. MCR score values and the error bars were calculated
from the average and the standard deviations obtained from undertaking MCR separately on the replicate
measurements.
All components showed higher scores (Figure 3) when the excitation and emission
polarizers had the same axis of polarization (VV and HH). This was expected due to the
slower fluorophore rotational speed in the more rigid protein. The normalized scores plots
show that each component changed at a different rate, which indicated different fluorophore
locations and processes at play. Furthermore, using the recovered pure components (Figure 2),
the anisotropy at each emission wavelength for each component was then calculated (Figure
corrections. DOI: 10.1016/j.aca.2015.06.011
.
Page 8 of 26
Figure 3: MCR scores plots and normalized MCR scores plots of the pure components extracted from HH (a/b),
VV (c/d), VH (e/f), and HV (g/h) polarized TSFS spectra of 1 mg mL
–1 HSA in PBS undergoing thermal
unfolding from 1070 °C and cooling from 7020 °C. MCR score values and the error bars were calculated
from the average and the standard deviations obtained from undertaking MCR separately on the replicate
measurements.
All components showed higher scores (Figure 3) when the excitation and emission
polarizers had the same axis of polarization (VV and HH). This was expected due to the
slower fluorophore rotational speed in the more rigid protein. The normalized scores plots
show that each component changed at a different rate, which indicated different fluorophore
locations and processes at play. Furthermore, using the recovered pure components (Figure 2),
the anisotropy at each emission wavelength for each component was then calculated (Figure