1 TC: 200 250 300 350 Wavelength (nm) UV/Vis spectrum of four brands of polysorbate 80, a common detergent used in protein purification. From Wuelfin, et al., "Polysorbate 80 UV/vis spectral and chromatographic characteristics-defining boundary conditions for use of the surfactant in dissolution analysis." 3. While the binding of Coomassie blue to proteins can lead to accurate measurements of protein concentration, there are situations where the absorbance values could be misleading. For example, detergents are often used in the purification of proteins. Certain detergents mixed in with the dye could lead to incorrect determinations of protein concentration. Take a look at the spectrum of the detergent polysorbate 80, above. If you had used this detergent to purify the protein in part c above, would you expect it to interfere with your concentration calculations? Why or why not? Absorbance (AU)
1 TC: 200 250 300 350 Wavelength (nm) UV/Vis spectrum of four brands of polysorbate 80, a common detergent used in protein purification. From Wuelfin, et al., "Polysorbate 80 UV/vis spectral and chromatographic characteristics-defining boundary conditions for use of the surfactant in dissolution analysis." 3. While the binding of Coomassie blue to proteins can lead to accurate measurements of protein concentration, there are situations where the absorbance values could be misleading. For example, detergents are often used in the purification of proteins. Certain detergents mixed in with the dye could lead to incorrect determinations of protein concentration. Take a look at the spectrum of the detergent polysorbate 80, above. If you had used this detergent to purify the protein in part c above, would you expect it to interfere with your concentration calculations? Why or why not? Absorbance (AU)
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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