cavette contain 2 ml substrate (catechol)and 0.1 ml enzyme extract and 0.9 ml of 0.2 M sodium acetate buffer
for blank-2ml catechol and 1 ml of sodium acetate buffer.Please give me easy explanations with formulas.Thanks u
Hi there,
plot A=f(t)
determine the laps of time during which variation of aborbance is linear
slope of the curve during this laps of time gives you the rate of the reaction
As you added 0.1mL of enzyme solution in the assay mixture, activity will be 10*inital rate/mL. If you know the protein content of your enzyme solution then you can calculate the activity per mg of protein.
The rest is about unit conversion. The rate is initially expressed as variation of OD per minute, if you know the molar extinction coefficient of the absorbing molecule you can convert it into mol/L/min and as the volume is 3mL you can calculate the rate as mol/min.
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