Instrumentation

Autoclave (10sq ft ) for contained biohazard waste processing.

From Left to right: a -80 freezer, a -20C freezer, and a second -20C freezer

The MicroCal PEAQ-ITC is a highly sensitive, low volume isothermal titration calorimeter.

Malvern Panalytical

Applications

1. Characterize interactions between proteins, nucleic acids, lipids, drugs and inhibitors.
2. Stoichiometry & thermodynamic parameters.
3. Correlate binding and activity.
4. Structure-activity relationships.
5. Hit validation, lead optimization, mechanism of action

Sample guidelines

• Both the cell and syringe samples should be in identical buffers. 
• Required volumes per measurement: 280ul of sample for the cell and 80ul of sample for the syringe.

Suggested concentrations

In the cell - at least 10x -20X the expected Kd, 350 ul per run, 280 ul will be injected.

In the syringe (ligand) - 10x above cell sample concentration, 80 ul/run

For example, for a 1 uM Kd a good titration will be -  20uM cell sample (350ul) and 200uM ligand (80 ul), any other higher concentrations with ratio 1:10 will work too.

All users will bring ultrapure methanol (~80 ml/all day run) and Pipet/tips/tubes for 80 ul ligand dispensing. HTS staff will train students/postdocs on ITC usage and enable access to the lab for the duration of the experiment. The users will sign into the log book and will also follow/log rinse and clean protocols during the experiment and at the end of the day. All issues encountered during the ITC runs will be entered into the log book.