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Analysis Interactions of Copper and Lysozyme by Isothermal Titration Calorimetry (ITC) (CAT#: STEM-MB-2551-LGZ)

Introduction

Although copper is essential to all living organisms, its potential toxicity to human health has attracted widespread concern. Previous studies have reported that copper can alter the physical properties of lysozyme. The direct combination of copper and lysozyme may cause changes in the conformation and function of lysozyme, thereby affecting the body's resistance to bacterial attack. Using isothermal titration calorimetry (ITC) to study the interaction of copper with lysozyme can provide a better understanding of the potential toxicity and toxicity mechanism of copper.




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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