Absorbance at 280, 545, 565, 620 and 650 nm
and absorption spectra were recorded on a
Perkin–Elmer (Beaconsfield, UK) Lambda-16
UV-Visible spectrophotometer with a 1 cm light
path. Fluorescence emission spectra were recorded
on a Shimadzu (Kyoto, Japan) RF5001 spectrofluorometer.
For steady state polarized fluorescence
measurements the spectrofluorometer was equipped with polarizers in the excitation and
emission paths. The anisotropy was calculated as
A=(IVV−GIVH)/(IVV+2GIHH), where G=IHV/
IHH, a correction factor for the polarization due
to the optics in the instrument. Protein concentrations
were chosen so that re-absorption of the
emission was negligible. All spectra were recorded
at room temperature.
The amounts of B-PE, R-PC and APC in the
different extracts and biliprotein containing solutions
were calculated from measurements of the
absorbance at 565, 620 and 650 nm using the
following equations.
Absorbance at 280, 545, 565, 620 and 650 nm
and absorption spectra were recorded on a
Perkin–Elmer (Beaconsfield, UK) Lambda-16
UV-Visible spectrophotometer with a 1 cm light
path. Fluorescence emission spectra were recorded
on a Shimadzu (Kyoto, Japan) RF5001 spectrofluorometer.
For steady state polarized fluorescence
measurements the spectrofluorometer was equipped with polarizers in the excitation and
emission paths. The anisotropy was calculated as
A=(IVV−GIVH)/(IVV+2GIHH), where G=IHV/
IHH, a correction factor for the polarization due
to the optics in the instrument. Protein concentrations
were chosen so that re-absorption of the
emission was negligible. All spectra were recorded
at room temperature.
The amounts of B-PE, R-PC and APC in the
different extracts and biliprotein containing solutions
were calculated from measurements of the
absorbance at 565, 620 and 650 nm using the
following equations.
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