Learning to use the ITC over the past week and a half has been a long process. At the beginning of the week, I got some pretty terrible results. Here's a small selection from the many failures I encountered.
Over time though, the results slowly got better. The goal is to have a smooth "hill" formed by the injection peaks which returns to a flat baseline about halfway to two-thirds of the way through the experiment. Here's a few my good data sets, which I was able to collect some good numbers from.
I was able to gather four pieces of data from this. The first is the binding energy, Kd. This is a measure of how strong the bond between two chemicals is. The next important number is the stoichiometry of the reaction, n. This represents the ratio of chemicals needed for the reaction to take place. Third, we have the change in enthalpy, delta H. This value is related to both the change in temperature and pressure/volume during the reaction. The last constant is the change in entropy, delta S. This is more or less a measure of the increase in disorder caused by the reaction.
The following graphs were obtained by plotting the area under each injection peak versus its peak number, and then fitting a curve to the data points. This gave us some very consistent results which were also pretty accurate. This is a good sign for future tests!
No comments:
Post a Comment