We read the absorbance value from the reaction. How can we interpret the absorbance value of the reaction into umol Pi/mg protein/hour?. Is there any formula to calculate the value?
Thank you.
Make a phosphate standard curve under identical buffer conditions as for the enzyme assay using a range of phosphate concentrations that includes the range generated by the reaction. Use that to convert absorbance to phosphate concentration (e.g. in µmoles/liter). Multiply by the reaction volume in liters, divide by the number of mg of protein in the reaction, and by the reaction time in hours.
I agree with Adam. Also, the standard curve should give you in which concentration range your assay delivers a linear relationship between phosphate concentration and absorbance. After making the standard curve you could adjust your assays to always run one standard phosphate concentration from the linear range of the curve.
Alia Aznan
I am also trying to estimate Na/K ATPase enzyme in tissue of rat but not getting results. I have followed method described by Tsimaratos et al 2001.
Can you forward the method adopted by you.
Regards
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