I would like to determine soil amylase activity at different intervals (5,10,20,30,45) from absorbance (of standard curve).
You must convert a change in absorbance to a concentration of amylase reaction product using a standard curve of product concentration v. absorbance. The initial rate of increase of product concentration (µmoles/minute), divided by the amount of protein (mg) in the reaction, is the specific activity.
You must convert a change in absorbance to a concentration of amylase reaction product using a standard curve of product concentration v. absorbance. The initial rate of increase of product concentration (µmoles/minute), divided by the amount of protein (mg) in the reaction, is the specific activity.
incubate the reaction mixture for such time intervals and take the readings. Thats it. And as said by Adam sir convert the absorbance with respect to standard curve. Important thing here is to have activity calculated in µmoles of product formed "per minute" for all the time intervals.
why diving on minutes???
i want 5minute and 10 , 20 , 30 , 45 continuously ??
Activity is mentioned in micromoles of product released per min. So for each time interval you gotta divide to know what is the reaction rate per min.
It is crucial that you measure the initial rate of reaction, before the substrate becomes deplete and the reaction slows down. If you make a graph of product formed (y-axis) versus time (x-axis), you can calculate the reaction rate as the slope of the line from the origin through the earliest data points before the graph starts to curve downwards.
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