The concentration of APC in solution can be determined by substituting the molecular weight, extinction coefficient and λ max into a derived form of the Beer - Lambert Law. A substance's λmax is the wavelength at which it experiences the strongest absorbance. For APC, this wavelength is 280 nm.
The absorbance at λmax can be measured using a spectrophotometer. There are some important things to keep in mind when measuring 280 absorbance:
The APC should be well-dissolved in solution. APC precipitation will cause inaccuracies in concentration calculations.
The absorbance reading should not exceed the maximum detection limit of the instrumentation. This can be identified by a plateau in the absorbance spectrum. If the absorbance spectrum plateaus, dilute the sample and try again.
This calculator assumes a 1 cm pathlength for instrumentation. Standard cuvettes for spectrophotometers are typically 1 cm. If a different pathlength is used, value should be corrected during data entry.
Protein
APC - Allophycocyanin
Absorbance at λmax
Dilution factor (optional)
Pathlength
cm
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