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When the concentration of a given protein is known, the concentration and molecular weight can be used to determine the molar extinction coefficient.
It is determined using the Beer-Lambert law: A = ε ∙ L ∙ c. A is the absorbance at 280 nm, L is the light path (cm), c is protein concentration (g/L), and ε is the extinction coefficient (L g-1cm-1). When the light path L = 1cm, ε = A/c. ε value corresponds to the slope of the line plotted between absorbance and concentration.
You then make the calculation of the extinction coefficient am (L mol-1cm-1) using the formula am = ε∙ MW, where MW is the molecular weight of the protein.
What we need from you to perform this analysis:
If you do not provide the buffer along with the sample we will use PBS as a standard buffer system.
