Rnase-2'Cmp Experiment Tutorial - MicroCal VP-ITC User Manual

4.3 Rnase-2'CMP Experiment Tutorial

Included with your instrument shipment, you should have received a test kit which includes
solutions of buffer, the ligand 2'CMP and the enzyme Ribonuclease A. If you have not received
this please contact info@microcalorimetry.com with shipping information and the sample will be
sent to you. Before your instrument was shipped from MicroCal, we used aliquots of the same
samples to generate a binding isotherm and have provided you with a copy of the results we
obtained, including the run parameters and the fitting parameters obtained from data analysis in
Origin. If your techniques are good, you should be able now to generate the RNase/2'CMP
binding isotherm and obtain fitting parameters very similar to those obtained at MicroCal with
your instrument.
The run parameters used in this experiment will be obtained from the rnase.inj run parameters
file. This experiment should follow all recommended system preparation, sample preparation,
loading and related operational standards outlined previously in this manual.
You should expect your results to be close to the data set sent with the samples. The following
percentage error data would be considered acceptable for a new user.
If everything has gone well to this point, then you should be ready to begin studies on your own
samples. Good luck!!
N (Stoichiometry)
Kb (binding Constant)
∆H (Enthalpy Change)
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Section 4: Experimental Tutorials
±5%
±15%
±5%