Analysis Interactions of Hyaluronidase (Hyal) and Epigallocatechin Gallate (EGCG) by Microscale Thermophoresis (MST) (CAT#: STEM-MB-2443-LGZ)

Introduction

Studying the binding affinity between biomacromolecules (such as enzymes) and small molecules (molecular weight less than 1000 Da) is an extremely challenging task because it is difficult to detect small changes caused by binding when using traditional techniques. Microscale Thermophoresis (MST) is a sensitive novel biophysical tool for the rapid and accurate determination of the dissociation constant (Kd) of enzyme-small molecule systems

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Principle Applications Procedure Materials Notes Related Products

Principle

One of the interacting molecules (mostly proteins) is labeled with fluorescent dye or combined with GFP label. The labeled protein and ligand molecules are placed in the capillary according to a specific concentration gradient. Infrared laser heating generates a microscopic temperature gradient field to undergo thermophoresis, with which the hydration layer, molecular size, electric charge and other molecular properties will change. Then the fluorescence distribution in the reaction system changes. In addition to accurately detecting interactions between biomolecules, MST can also obtain other parameters related to interactions between biomolecules by calculating dissociation constants (Ks), and achieve accurate qualitative analysis.

Applications

For characterizing thermodynamic parameters of biomolecular interactions.

Procedure

1. Sample processing.
2. MST detection.
3. Data analysis.

Materials

• Sample Type: protein 10uM/ 100ul or 50 micrograms, small molecules 100uM/200ul, protein peptides need to be sent at low temperature

Notes

In order to ensure the reliability of the test, some samples should be provided as much as possible.

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