Assessing the Specificity of Monoclonal Antibody 8E7 for β-Catenin
Author Information
Author(s): Meghan T. Maher, Annette S. Flozak, Alyssa M. Hartsell, Susan Russell, Rohinee Beri, Ofra N. Peled, Cara J. Gottardi
Primary Institution: Northwestern University Feinberg School of Medicine
Hypothesis
Does monoclonal antibody 8E7 specifically recognize the unphosphorylated form of β-catenin without cross-reacting with other nuclear antigens?
Conclusion
Monoclonal antibody 8E7 can cross-react with a non-specific nuclear antigen, which complicates the interpretation of β-catenin signaling.
Supporting Evidence
- The 8E7 antibody cross-reacts with a nuclear antigen that is not β-catenin.
- Using milder antigen retrieval methods can reduce non-specific staining.
- Cadherin overexpression can help confirm the specific detection of β-catenin.
Takeaway
This study shows that an antibody used to detect a protein called β-catenin might also stick to other things in cells, making it hard to tell what's really happening.
Methodology
The study involved immunofluorescence and immunoblotting techniques to assess the specificity of the 8E7 antibody in various cell lines.
Potential Biases
Potential bias in interpreting results due to non-specific staining by the antibody.
Limitations
The study primarily focuses on specific cell lines, which may not represent all biological contexts.
Digital Object Identifier (DOI)
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