8
Measure Proteins and Labels
78
NanoDrop One User Guide
Thermo Scientific
Related Topics
Dye concentrations are calculated from the absorbance
value at the dye’s analysis wavelength, the dye’s
extinction coefficient, and the sample pathlength. A
sloped-line dye correction may also be used.
Dye absorbance
• Dye absorbance values are measured at specific wavelengths. See
for analysis wavelengths used.
• If Sloping Dye Correction is selected, a linear baseline is drawn
between 400 nm and 750 nm and, for each dye, the absorbance value
of the sloping baseline is subtracted from the absorbance value at each
dye’s analysis wavelength. Baseline-corrected dye absorbance values are
reported and used to calculate dye concentrations.
Dye correction
• Pre-defined dyes have known correction values for A260 and A280.
See
• A280 dye correction is subtracted from
used to
calculate protein concentration.
Sample Pathlength
• For micro-volume measurements, the software selects the optimal
pathlength (between 1.0 mm and 0.03 mm) based on sample
absorbance at the analysis wavelength.
• For cuvette measurements, pathlength is determined by the cuvette
Pathlength setting in the software (see
• Displayed spectra and absorbance values are normalized to a 10 mm
pathlength equivalent.
Reported Values
•
Protein concentration.
Reported in selected unit (mg/mL or μg/mL).
Calculations are based on Beer-Lambert equation using corrected
protein absorbance value.
•
Dye1/Dye2 concentration
. Reported in pmol/μL. Calculations are
based on Beer’s Law equation using (sloping) baseline-corrected dye
absorbance value(s).
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