I already have plotted a maltose standard curve but the readings for absorbance of amylase activity assay are beyond the range of the maltose standard curve. The abosrbance reading for 1.8mg of maltose is 0.37 whereas the absorbance readings for the amylase activity assay all start from 1.0 absorbance unts.
Dear Akshay
There is some problem with your assay. As I had done it 5 years ago and I checked the data and I am attaching the excel sheet for your kind information. I have assayed using 5 mg/ml maltose.
Further you have to dilute your enzyme so that absorbance reading should not go beyond detection limits and then calculate the unit/ml by multiplying with dilution factor.
You may go through out paper on Comparative fermentation studies for amaylase production in my profile.
Dear Akshay
firstly the absorbance must be between 0.1 and 0.9 to be accurate, so you may need to dilute your samples
and after you plot standard curve of maltose you can use the line equation to find any unknown sample.
regards
Dear Akshay,
You should pay some attention to your blank tube. Maybe a better blank missing only the maltose could be enough to move your readings to the correct range as long as your equipment allows for that.
Hi, I agree. The absorbance lectures should be 0.1 up to 1.2 using a maltose concentration between 0 to 25mM. You need assay different enzyme concentrations in order to get absorbance in the equation range.
You may have to dilute your enzyme if you noticed the concentration is too high, take note of the dilution you used so that you will use the dilution factor to calculate your enzyme activity.
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