Which Antibody Is Used for Immunoprecipitation in Protein Ubiquitination
Anti-ubiquitin antibodies serve as vital tools for investigating protein ubiquitination. They are particularly valuable in immunoprecipitation assays for the enrichment and detection of ubiquitinated proteins. This paper presents a detailed immunoprecipitation protocol using anti-ubiquitin antibodies aimed at exploring protein ubiquitination processes, essential for understanding mechanisms such as protein degradation and signal transduction.
Materials and Reagents
1. Anti-ubiquitin antibody
2. Target protein-specific antibody
3. Protein A or G magnetic beads
4. Immunoprecipitation buffer
5. Lysis buffer
Methodology
1. Isolate total proteins from cell or tissue samples.
2. Conduct immunoprecipitation using anti-ubiquitin antibodies coupled with protein A or G magnetic beads.
3. Wash the magnetic beads with immunoprecipitation buffer to eliminate non-specific protein interactions.
4. Elute proteins specifically bound to the beads using an appropriate elution buffer.
5. Subject the eluted proteins to SDS-PAGE followed by immunoblotting to identify ubiquitinated proteins.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
Related Services
How to order?
