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Analysis Interactions of Biomolecule–nanoparticle by Isothermal Titration Calorimetry (ITC) (CAT#: STEM-MB-2524-LGZ)

Introduction

Nanomaterials (NMs) are frequently exposed to a wide range of biomolecules in varying abundances. Biomolecular adsorption driven by various interfacial forces determines the surface structure and composition of NMs, which in turn determines their functionality and reactivity of the adsorbed biomolecules. Isothermal titration calorimetry (ITC) is a non-destructive technique to quantify thermodynamic parameters by in situ measurement of heat absorption or release associated with interactions.




Principle

A reactant is placed in a temperature-controlled sample cell and coupled to the reference cell through a thermocouple loop. The sample cell and the reference cell are in the same external environment. A specific titration agent (selected as required by the test) is added to the sample cell quantitatively. The change in energy in response can be sensitively detected and triggered by a positive or negative feedback thermostat to keep the temperature constant.

Applications

For characterizing thermodynamic parameters of biomolecular interactions.

Procedure

1. Preparation before the experiment.
2. Add 40ul of ligand or small molecule protein solution to the titration syringe, 200ul of large molecule protein solution to the sample pool, and 200ul of distilled water to the reference pool.
3. Set Experimental Parameters in the interface.
4. In the set up interface, set the storage path of experimental data and experimental methods, as well as the current user.
5. Put the titration syringe into the sample pool and click Start to start the experiment. Display real-time images of the experiment on the Real Time Plot interface.

Materials

• Sample Type: protein 50 micrograms, small molecules 10mM/500ul, the higher the sample concentration is the better, protein peptides need to be sent at low temperature.
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