TUBA1 Antibody
Description
TUBA1A Protein and Its Biological Significance
TUBA1A (Tubulin Alpha-1A Chain) is encoded by the TUBA1A gene and constitutes the predominant α-tubulin isoform in the developing human brain. It forms heterodimers with β-tubulin to assemble microtubules, which are vital for:
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Neuronal migration and cortical layering during brain development .
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Intracellular transport, cell division, and cytoskeletal integrity .
Mutations in TUBA1A disrupt microtubule function, leading to tubulinopathies such as lissencephaly, microcephaly, and intellectual disability .
Development and Validation of TUBA1A Antibodies
TUBA1A antibodies are designed to distinguish TUBA1A from other α-tubulin isoforms (e.g., TUBA1B, TUBA1C) due to high sequence homology. Key features include:
Examples of Commercial Antibodies:
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ab95966 (Abcam): Rabbit polyclonal antibody validated in WB and IF, detecting a 50 kDa band .
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MACO0009 (Assay Genie): Mouse monoclonal antibody for human samples, optimized for WB and IP .
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YL1/2 (Thermo Fisher): Binds tyrosylated α-tubulin, serving as a loading control .
3.1. Neurodevelopmental Studies
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Neurite Outgrowth: TUBA1A knockdown in mice impairs neurite extension and growth cone dynamics, linked to MAP1B mislocalization .
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Commissure Defects: Tuba1a<sup>N102D</sup> mutants exhibit normal cortical layering but fail to form axon commissures .
3.2. Disease Mechanisms
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Tubulinopathies: Antibodies identify pathogenic mutations (e.g., R402C, L397P) that disrupt tubulin folding or microtubule assembly .
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Apoptosis: Codon-modified Tuba1a alleles increase cortical apoptosis, correlating with severe neurodevelopmental phenotypes .
3.3. Technical Advancements
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Tagging Methods: Novel TUBA1A-His6 tools enable live visualization of microtubule incorporation without functional impairment .
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Post-Translational Modifications: Antibodies detect detyrosination or polyglutamylation, critical for neuronal microtubule specialization .
Comparative Analysis of TUBA1A Isoforms
TUBA1A shares high homology with other α-tubulins but has unique functional domains:
Challenges and Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- These findings, along with the three-dimensional location of the phosphorylation site, suggest that microtubules undergo depolymerization in response to hyperosmotic stress through alpha-tubulin phosphorylation. PMID: 23628996
Q&A
TUBA1A Antibody: Academic Research FAQs
Advanced Research Questions
How can I resolve discrepancies in TUBA1A antibody specificity across isoforms?
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Challenge: TUBA1A shares >90% sequence homology with other α-tubulin isoforms (e.g., TUBA3, TUBA1B) .
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Solutions:
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Knockdown Validation: Use siRNA targeting TUBA1A in HEK-293T cells to confirm band loss at ~50 kDa .
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Post-Translational Modification (PTM) Analysis: Employ antibodies targeting PTM-specific epitopes (e.g., acetyl-K40 in ab95966 ).
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Cross-Reactivity Testing: Compare reactivity in lysates from organisms lacking TUBA1A (e.g., Drosophila melanogaster ).
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What methodologies validate TUBA1A’s role in neuronal development using antibody-based assays?
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Functional Assays:
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Data Interpretation:
Methodological Troubleshooting
How should I address non-specific binding in TUBA1A immunohistochemistry (IHC)?
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Blocking: Use 5% BSA with 0.3% Triton-X 100 to reduce background .
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Antigen Retrieval: Optimize heat-mediated retrieval (pH 6.0 citrate buffer) for formalin-fixed brain tissues .
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Validation Table:
Data Contradiction Analysis
Why do TUBA1A expression levels vary between qPCR and Western blot data?
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Transcript vs. Protein Stability: mRNA levels (e.g., TNFRSF12A in scleroderma ) may not correlate with protein due to translational regulation.
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Antibody Sensitivity: Compare multiple antibodies (e.g., ab95966 vs. A03989) and confirm with mass spectrometry .
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Normalization Errors: Re-express data relative to both GAPDH (mRNA ) and β-actin (protein ).
