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An antibody against the Alzheimer's disease amyloid precursor protein recognizes distinct conformational isoforms
P Piccardo1, A Dagenais, A C Cuello
1Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec, Canada.
Abstract:
The Alzheimer's disease amyloid precursor protein (APP) consists of several isoforms, which are extensively post-translationally modified and processed. A monoclonal antibody, MAbE1, was raised against a synthetic peptide from an extracellular domain that is common to all isoforms of APP. Immunoblots and immunolocalization studies on cells of neuronal and other origins demonstrated that this antibody recognized a subclass of APP isoforms when compared to a monoclonal antibody raised against a bacterial fusion protein of APP, MAb22C11. Prominent protein bands of 71 kDa and 120 kDa were only detected on immunoblots of cell lysates and no immunoreactivity was observed in protein samples obtained from cell conditioned media. Immunofluorescence labelling with MAbE1 revealed predominantly perinuclear staining of cells of neuronal and glial origin. The data suggest that this monoclonal antibody detects distinct conformational isoforms of APP present in intracellular compartments.
Insights
A new antibody, MAbE1, identifies specific Alzheimer's disease amyloid precursor protein (APP) isoforms. These distinct APP forms are primarily located within intracellular compartments of neuronal and glial cells.
Area of Science:
- Neuroscience
- Molecular Biology
- Immunology
Background:
- Alzheimer's disease is linked to the amyloid precursor protein (APP).
- APP exists in multiple isoforms with extensive post-translational modifications.
- Understanding APP isoform localization is crucial for Alzheimer's research.
Purpose of the Study:
- To develop and characterize a novel monoclonal antibody (MAbE1) targeting specific APP isoforms.
- To compare the reactivity of MAbE1 with a standard APP antibody (MAb22C11).
- To investigate the cellular localization of APP isoforms recognized by MAbE1.
Main Methods:
- Generation of MAbE1 against a synthetic APP peptide.
- Immunoblot analysis of cell lysates and conditioned media.
- Immunofluorescence microscopy for cellular localization studies.
- Comparison with MAb22C11.
Main Results:
- MAbE1 recognized a subclass of APP isoforms, distinct from MAb22C11.
- Specific protein bands of 71 kDa and 120 kDa were detected in cell lysates.
- No immunoreactivity was found in cell conditioned media.
- MAbE1 showed predominantly perinuclear staining in neuronal and glial cells.
Conclusions:
- MAbE1 identifies distinct conformational APP isoforms.
- These specific APP isoforms are primarily located in intracellular compartments.
- The findings provide insights into APP processing and localization in neuronal and glial cells.