Immunological Detection Method for Protein Acetylation Detection

Acetylation is a common type of post-translational modification of proteins, which plays an important role in regulating the function and stability of proteins. Immunodetection is a commonly used method for detecting protein acetylation, including Western blot and Immunoprecipitation.

 

1. Western Blot

The principle of Western blot is to bind the target protein with a specific antibody, and then detect the protein through fluorescence or chemiluminescence analysis. In the detection of acetylated proteins, the test sample needs to be run on SDS-PAGE electrophoresis first, then the proteins after electrophoresis are transferred to the PVDF membrane, followed by incubation with a specific antibody (usually rabbit anti-acetylated protein antibody), then incubation with the secondary antibody binding to the rabbit antibody, and finally detection is carried out through chemiluminescence reaction.

 

2. Immunoprecipitation

Immunoprecipitation uses a specific antibody to recognize and bind to the target protein, and then the protein and its binding material are enriched and separated by centrifugation. For the detection of acetylated proteins, immunoprecipitation can be used with anti-acetylated protein antibodies to enrich the acetylated proteins in the sample, and then detected by Western blot or quantified by mass spectrometry.