western blot
Definition
Western blot, also known as immunoblotting, is a widely-used analytical technique in proteomics that detects specific proteins in a complex biological sample. The method involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and probing with specific antibodies that bind to the target protein. Detection systems then visualize the antibody-protein complexes, allowing researchers to confirm protein presence, determine relative abundance, assess molecular weight, and evaluate post-translational modifications. Western blotting is essential for validating proteomic findings, confirming gene expression at the protein level, and studying protein regulation in various biological contexts including disease states and drug responses.
Visualize western blot in Nodes Bio
Researchers can integrate western blot validation data into Nodes Bio to confirm protein-protein interactions identified through mass spectrometry or co-immunoprecipitation studies. By mapping validated proteins as nodes and their confirmed interactions as edges, users can build high-confidence protein networks. Western blot quantification data can be visualized as node attributes, enabling comparison of protein expression levels across experimental conditions within pathway contexts.
Visualization Ideas:
- Protein-protein interaction networks with western blot-validated nodes highlighted
- Signaling pathway maps showing phosphorylation states confirmed by western blot
- Comparative protein expression networks across experimental conditions with quantified western blot data
Example Use Case
A cancer research team investigating resistance to EGFR inhibitors performs western blots to validate mass spectrometry findings. They detect elevated phosphorylation of AKT and ERK proteins in resistant cell lines compared to sensitive ones. By confirming these key signaling proteins through western blot, they validate their proteomic network analysis showing activation of bypass signaling pathways. This validated data helps prioritize combination therapy targets, leading to testing dual EGFR-AKT inhibition strategies in preclinical models.