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Binding of a small molecule, compound B, to a protein labeled with Alexa Fluor 647 was measured by MicroScale Thermophoresis at 10% excitation power and
Binding of a small molecule, compound B, to a protein labeled with Alexa Fluor 647 was measured by MicroScale Thermophoresis at 10% excitation power and medium MST power. 2nM fluorescent protein was mixed 1:1 with a serial dilution of compound B from 1.3mM and plotted below, then fit using the quadratic form of the binding isotherm. From the graph below, can you estimate the KD of this interaction? Explain what experimental parameters allow or don't allow you to make a reasonable estimate. Then make your best guess of the KD and explain your results. Binding of a small molecule, compound B, to a protein labeled with Alexa Fluor 647 was measured by MicroScale Thermophoresis at 10% excitation power and medium MST power. 2nM fluorescent protein was mixed 1:1 with a serial dilution of compound B from 1.3mM and plotted below, then fit using the quadratic form of the binding isotherm. From the graph below, can you estimate the KD of this interaction? Explain what experimental parameters allow or don't allow you to make a reasonable estimate. Then make your best guess of the KD and explain your results
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