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Chapter 19: Q21P (page 488)
What is the advantage of a time-resolved emission measurement with Eu3+versus measurement of fluorescence from organic chromophores?
The main advantage is that they are plagued by background fluorescence from solvent, solutes, and particles.
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Scatchard plot for binding of estradiol to albumin. Data in the table come from a student experiment to measure the binding constant of the radioactively labeled hormone estradiol (X)to the protein, bovine serum albumin was equilibrated with various concentrations of albumin for A small fraction of unbound estradiol was removed by solid phase microextraction () and measured by liquid scintillation counting. Albumin is present in large excess, so its concentration in any given solution is essentially equal to its initial concentration in that solution. Call the initial concentration of estradiol and the final concentration of unbound estradiol [X]. Then bound estradiol isand the equilibrium constant is
which you can rearrange to
localid="1663648487221"
A graph of versus [P]should be a straight line with a slope of K.The quotient is equal to the counts of radioactive estradiol extracted from a solution without albumin divided by the counts of estradiol extracted from a solution with estradiol. (b) What fraction of estradiol is bound to albumin at the first and last points?
Infrared spectra are customarily recorded on a transmittance scale so that weak and strong bands can be displayed on the same scale. The region near in the infrared spectra of compounds A and B is shown in the figure. Note that absorption corresponds to a downward peak on this scale. The spectra were recorded from a 0.0100M solution of each, in cells with 0.00500 - cm path lengths. A mixture of A and B in a 0.00500 - cm cell gave a transmittance 34.0 % of at2022cm and 383% at 1093 cm. Find the concentrations of A and B.
A study was conducted with derivatives of the DNA nucleotide bases adenine and thymine bound inside micelles () in aqueous solution.
Sodium dodecyl sulfate forms micelles with the hydrocarbon tails pointed inward and ionic headgroups exposed to water. It was hypothesized that the bases would form ahydrogen-bonded complex inside the micelle as they do in DNA:
To test the hypothesis, aliquots of 5.0 mMadenine derivative were mixed with aliquots of 5.0 mMthymine derivative in proportions shown in the table. Each solution also contained 20mMsodium dodecyl sulfate. The concentration of product measured by nuclear magnetic resonance also is shown in the table. Are the results consistent with formation of a 1:1complex? Explain your answer.
Figure 19-6 is a Scatchard plot for the addition of 0-20nM antigen X to a fixed concentration of antibodyPrepare a Scatchard plot from the data in the table and find K for the reaction. The table gives measured concentrations of unbound X and the complex PX. It is recommended that the fraction of saturation should span the range ,-0.2-0.8. What is the range of the fraction of saturation for the data?
Find the concentration of [X] if the absorbance are 0.700 at 272 nm and
0.550 at 327 nm.
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