Dear Everyone,
I'm doing experiment on finding enzyme activity and specific activity with different concentration of enzyme and same concentration of substrate. The enzyme comes from blood plasma. The undiluted concentration of the blood plasma is 48.208 mg/mL and the blood plasma was diluted with multiple dilution rate (1/4,1/8,1/16,1/32,1/64,1/128). Afterwards, I managed to calculate the amount of blood plasma in each dilution and gained the reaction rate in each different concentration. How do I find the enzyme activity and specific activity from this?
The enzyme activity is mostly represented as micromoles/min/ml and specific activity as micromoles/min/mg protein.
You need to identify protein concentration by Folin-lowry /bradford method/biuret method. For blood plasma/serum, preferrable is biuret method due to higher amount of proteins.
https://www4.uwsp.edu/chemistry/tzamis/365f00pdfs/specificactivity10.pdf
https://en.wikipedia.org/wiki/Enzyme_assay
Hi there,
As the relation between reaction rate and amount of protein is linear, the slope is actually the specidfic activity expressed in µM/min/mg of protein. If you want to convert it into µmol/min/mg, you just have to multiply by the volume of the reaction expressed in L. As you know the protein concentration of your protein sample you can express activity in terms of µmol/min/mL (by dividing AS by protein concentration of your protein sample) which is AE.
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