Analysis Interactions of RNA–RNA by Microscale Thermophoresis (MST) (CAT#: STEM-MB-2473-LGZ)

Introduction

Microscale Thermophoresis (MST) is the directed movement of particles in a microscopic temperature gradient. Affinity is determined by measuring trace thermal changes resulting from changes in the hydration layer (usually caused by changes in the structure/conformation of biomolecules). Even small changes like protein phosphorylation or small molecules binding to a target can be measured. MST also allows measurement of intermolecular interactions directly in solution without the need for a fixed surface (no fixation required). Evaluation of RNA–RNA binding is crucial for in vitro studying of molecular mechanisms, for example, the interaction of noncoding RNAs (ncRNAs) with their respective targets.

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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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