Quantitative analysis of carbohydrates is a fundamental analytical tool used in many aspects of biology and chemistry. We have adapted a technique developed by Mathews et al. using an inexpensive scanner and open-source image analysis software to quantify amylase activity using both the breakdown of starch and the appearance of glucose. Breakdown of starch is analyzed using the iodide/iodine reagent, while the appearance of glucose is followed using Benedict's reagent. The progress of Benedict's reagent is quantified using the inherent color change of a blue solution into a red precipitate. The reaction velocity at different time points is then plotted and graphed using an Excel spreadsheet. This laboratory emphasizes that enzymes mediate the conversion of a substrate into a product and that either the concentration of product or reactant may be used to follow the course of a reaction.
This cell line, Hep293TT, is a valuable resource for the study of childhood liver cancer and may potentially provide a tool in the development of new agents.
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