my enzyme is thermophile and has two site binding Ca. I want to examine the role of these Ca in stability and activity. what is a range of concentration of EDTA used?
depend on the concentrations of your enzyme and the presence of other cations in your solutions. Start with 1, 2, 5 on to 10 mM and analyse the activities. Once you got the effective concentrations in reducing the activities, you can add back the Calcium and test again the activities and find the level of Calcium required for activity and stability. Sometimes the Calcium effect is also dependant on the type of anion used
Hi, I suggest to use concentrations of 0.1 mM to 5 mM. EDTA is a general chelating agent and if it decreases activity of your enzymes it may refer to removing ca from active site of enzyme. Also it may denature proteinaceous structure of the enzyme by removing ca.