I performed spectrophotometry on 6 cuvettes containing NAD+, Buffer, Ethanol, H2O, and ADH, with varying concentrations of ethanol.
I have the slopes of the 6 graphs I made from these UV specs. Now I need help turning these slopes into Vo values. In order to make the Michaelis-Menten graphs, I need to plot Vo values vs. [Ethanol]. I believe this will involve the Beer's equation, but I'm not sure how to proceed and I need help from someone with a good understanding of Michaelis-Menton plots. The wavelength used was 340nm, and the E340nm for NADH = 6220 M-1cm-1.
My slopes are as follows:
Cuvette 1: 0.004
Cuvette 2: 0.006
Cuvette 3: 0.008
Cuvette 4: 0.009
Cuvette 5: 0.001 *outlier
Cuvette 6: 0.010
The ethanol concentrations were as follows:
cuvette 1: 25 microliters
cuvette 2: 50 microliters
cuvette 3: 125 microliters
cuvette 4: 250 microliters
cuvette 5: 375 microliters
cuvette 6: 750 microliters
If you can help me to find the Vo values for my plot that would be great. Please let me know if I can provide anymore helpful information. Thank you.
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