Process of IP Mass Spectrometry

The process of IP-MS (Immunoprecipitation Tandem Mass Spectrometry Analysis) mainly includes the following steps.

 

Analysis Workflow

1. Sample Preparation

Prepare cell lysate or tissue lysate containing target proteins according to experimental requirements.

 

2. Immunocomplex Preparation

Combine the antibody with a solid carrier such as magnetic beads or agarose to form an immunocomplex.

 

3. Sample Processing

Add the immunocomplex into the sample, allowing it to bind with the target protein.

 

4. Immunoprecipitation

Use magnetic beads or agarose or other solid carriers to precipitate the immunocomplex, thereby enriching the target protein and its interacting proteins.

 

5. Washing

Wash the immunocomplex with washing buffer to remove non-specifically bound proteins.

 

6. Elution

Use elution buffer to elute the target protein and its interacting proteins from the immunocomplex.

 

7. Proteolytic Digestion

Digest the purified protein complex into peptides for subsequent mass spectrometry analysis.

 

8. Mass Spectrometry Analysis

Identify and quantify proteins by analyzing the mass-to-charge ratio (m/z) and mass spectrometry data of peptides.