here am attaching a file of readings ,Total rxn volume is 3 ml, enzyme volume is 0.1ml.
Hello Laita, no one knows what kind of calculations or determinations you need. Please make your question clear and articulate the problem area properly.
Hello, see attached file example of calculation for you.
Hope this info will help you.
Good luck!
Maurice Ekpenyong Sir I want to calculate the total antioxidant enzyme activity ( SOD) in plant tissue i.e Launea in FW, for that i carried out the NBT assay of SOD now i want to calculate the units of SOD (Unit/ml) i have attached the file of my readings , out of two sets of my plant sample one was in dark absorbance is 0.016 and other was in light absorbance is 0.310, and that of control is 0.589, Total rxn volume was 3 ml and that of volume used was 0.1ml.
First prepare a rate curve. The rate of reaction can be obtained by regression of the uninhibited linear portion of the rate curve. The equation for this linear regression will be: Y = aX + b, where a is the decomposition rate or (∆Abs560nm/min). A less preferred way to obtain the rate is to select two data points in the rate curve and to calculate rates according to the following equation: Rate (ΔAbs560nm/min) = (Y2 – Y1)/(X2 – X1).
Calculate average rates from duplicate or triplicate assays.
2. Obtain the ratio of sample rate and blank rate using the equation Ratio (s/b) = Rate (s)/Rate (b); where Rate (s) is the average auto-oxidation rate of a sample, and Rate (b) is the average auto-oxidation rate of the blank.
3. Calculate percentage for SOD inhibition of baseline (blank) reaction rate as: % Inhibition = (1 - Ratio s/b] * 100%
4. SOD activity measured using the classical “cytochrome c reduction” method is found to be approximately 1.25 times measurable percent inhibition of hematoxylin auto-oxidation as established through in-house trials.
The SOD concentration in classical McCord Fridovich “cytochrome c reduction” unit activity is therefore calculated as: [SOD U/mL]s = 1.25 * (% Inhibition). This value is representative of the SOD activity in the sample added to the reaction mixture not the final concentration in the reaction mixture.
5. The SOD concentration in the original sample (“cytochrome c reduction” U/mL) is calculated using the equation: [SOD U/mL]os = [SOD U/mL]s * (Sample Dilution Factor)
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