Protein Group Verification

Proteome validation aims to confirm the existence of proteins, identify their characteristics, and understand their functions and interactions in biological processes. This validation method is crucial for a deep understanding of the roles and functions of proteins in organisms, contributing to the advancement of biomedical research and drug development. Proteome validation usually includes the following steps.

 

Protein Isolation

The target protein needs to be separated from the biological sample. Techniques such as gel electrophoresis, high-performance liquid chromatography (HPLC), ultracentrifugation, etc., are used.

 

Immunoassay

A common proteome validation method is immunoassay, i.e., using specific antibodies to detect the target protein. This can be achieved through techniques like Western blot, ELISA (enzyme-linked immunosorbent assay), or immunoprecipitation. In these methods, antibodies specifically bind to the target protein, thus confirming its existence.

 

Mass Spectrometry Analysis

It is used to identify the amino acid sequence, mass, post-translational modifications, and interactions of the protein with other proteins. Common mass spectrometry techniques include tandem mass spectrometry (MS/MS), liquid chromatography-mass spectrometry (LC-MS), etc.

 

Functional Validation

This is achieved through in vitro laboratory techniques, cell culture experiments, or mouse models. For example, the biological function of the target protein can be studied by knocking out or overexpressing it.

 

Structural Analysis

For the structural validation of proteins, techniques such as X-ray crystallography or nuclear magnetic resonance (NMR) are typically used to decode the three-dimensional structure of the protein.