How do I convert from ΔA340 to (μmol/min)/ml enzyme?
I am working on a lab report for biochemistry, and we are asked to calculate the (μmol/min)/ml enzyme for a GOT activity assay. I asked my professor and he just told me to look at the previous lab where we did the same calculation (but I got it wrong then too). No one in the class knows how to do this calculation, and even the professor has told us that the text book is no good for help with calculations. The entire lab report hinges on this calculation so it is critical. Thanks for any help.
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What you are measuring is probably the change in absorbance at 340 nm for either a reactant or product of the enzyme catalyzed reaction. Assuming you have the extinction coefficient for the compound, it is a simple matter to convert to change in concentration. You would just use Beer's law, ΔA=EbΔc and solve for Δc. Multiplying by the total reaction volume gives you moles, which you convert to μmol. You then divide this by the time interval over which the change occurs and then divide by the volume of enzyme extract to get your activity.
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