NanoPhotometer
®
N120/NP80/N60/N50/C40 User Manual Version 4.3.1
79
extinction coefficient defined by the user. The protein concentration is calculated with or
without background correction as follows:
With background correction:
C =
A
280
A
* ε
prot
* Ð
Without background correction:
C = A
280
* ε
prot
* Ð
C Concentration (mg/ml)
A
280
Absorbance at 280 nm (10 mm path) or entered wavelength
A
BKG
Absorbance at selected background wavelength (10 mm path)
ε
prot
Extinction coefficient/protein factor in g*cm/l
Ð Dilution factor
Table 3. Protein extinction coefficients (ε
prot
)
Mol. Ext. Coeff. [M
-1
*cm
-1
]
Dye-labeled Protein UV280 Concentration
For dye-labeled proteins, the concentration of the protein is calculated using a modified form
of the Beer-Lambert equation. For these calculations, the instrument considers the
absorption maximum of the dye, and a certain dye-specific correction factor at 280 nm
(see Table 4 on page 82). The dye concentration is calculated with or without
background/dye correction for are as follows:
With background and with dye correction:
C =
A
280
A
BKG
cf
dye
*
A
max,dye
A
BKG
* ε
prot
* Ð
With background and without dye correction:
C = (A
280
A
BKG
) * ε
prot
* Ð
Without background and with dye correction:
C =
A
280
cf
dye
* A
max,dye
* ε
prot
* Ð
Without background and without dye correction:
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