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The Bradford protein assay ... Compute the extinction coefficient and calculate the concentrations of the unknown samples. Procedure (Micro Assay, 1-10 μg protein/mL)
In biochemistry, the molar absorption coefficient of a protein at 280 nm depends almost exclusively on the number of aromatic residues, particularly tryptophan, and can be predicted from the sequence of amino acids. [6] Similarly, the molar absorption coefficient of nucleic acids at 260 nm can be predicted given the nucleotide sequence.
Extinction coefficient refers to several different measures of the absorption of light in a medium: Attenuation coefficient , sometimes called "extinction coefficient" in meteorology or climatology Mass extinction coefficient , how strongly a substance absorbs light at a given wavelength, per mass density
At a wavelength of 260 nm, the average extinction coefficient for double-stranded DNA is 0.020 (μg/mL) −1 cm −1, for single-stranded DNA it is 0.027 (μg/mL) −1 cm −1, for single-stranded RNA it is 0.025 (μg/mL) −1 cm −1 and for short single-stranded oligonucleotides it is dependent on the length and base composition.
Once a reference signal is acquired, it is compared to the transmitted light Is of the measured sample. In order to calculate the associated extinction coefficient μ t, Beer's law can be applied as follows: I s = I o exp(-μ t d), where d is the sample thickness. The extinction coefficient is therefore: μ t = -ln(I s /I o)/d.
Thus a scattering coefficient μ s and an absorption coefficient μ a can be combined into a total extinction coefficient μ = μ s + μ a. [ 6 ] Importantly, Beer also seems to have conceptualized his result in terms of a given thickness' opacity, writing "If λ is the coefficient (fraction) of diminution, then this coefficient (fraction) will ...
The attenuation coefficient of a volume, denoted μ, is defined as [6] =, where Φ e is the radiant flux;; z is the path length of the beam.; Note that for an attenuation coefficient which does not vary with z, this equation is solved along a line from =0 to as:
The SI unit of mass attenuation coefficient is the square metre per kilogram (m 2 /kg). Other common units include cm 2 /g (the most common unit for X-ray mass attenuation coefficients) and L⋅g −1 ⋅cm −1 (sometimes used in solution chemistry). Mass extinction coefficient is an old term for this quantity. [1]
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