Question

Annabeth is testing the effect of substrate concentration on enzyme activity using cellobiase and p-nitrophenyl glucopyranoside. After she adds STOP solution to her reactions and measures the absorbances, she discovers that one sample has an absorbance that is beyond the range of her standard curve. How can Annabeth determine the concentration of product for this sample?

Answer 1

Annabeth can do the following

1. Single out the sample which is showing a high absororbance

2. Prepare serial dilutions of the sample upto 10^-3, but some times more dilutions may be required

3. Measure the absorption of the diluted samples

4. Select the absorption than can fit the range, let it be sample A of dilution 10^-a

5. Plot and calculate the contration of sample A

6. Multiply the concentration by dilution factor (10^a) to get the original concentration of the sample