Live Cell Protein Interaction Thermal Stability Detection

Thermal stability detection of live cell protein interaction is an experimental method used to study the interaction between proteins and their stability under different temperature conditions. This technique is particularly important in biomedical research as it can help understand the function of proteins and their role in diseases. The following are detailed steps of this technique:

 

1. Cell Cultivation

First, cells containing the protein of interest need to be cultivated. These cells can be cells naturally expressing the target protein, or they can be genetically modified to express a specific protein.

 

2. Heat Treatment

The cells are treated at different temperatures for a certain period. The purpose of this step is to induce structural changes in the protein, including dissociation and denaturation. The choice of temperature depends on the specific needs of the research.

 

3. Cell Lysis

After heat treatment, cells are lysed to release their intracellular proteins. This is usually achieved by adding lysis buffer and physical methods (such as ultrasonic treatment).

 

4. Separation and Purification

Next, centrifugation and other methods are used to separate out the undenatured, still stable proteins. This step can be accomplished by collecting the supernatant, as denatured proteins typically precipitate.

 

5. Protein Analysis

Finally, Western Blot, mass spectrometry or other biochemical methods are used to analyze these proteins. By comparing samples treated at different temperatures, researchers can determine the thermal stability of the proteins and their interactions with each other.

 

This method can reveal how proteins interact and change under disease conditions or during drug treatment, and it has immense value for drug discovery and biomedical research. Through this method, scientists can better understand the function of proteins and how they interact with each other within cells.