TUBA1A Antibody
Product Specs
Target Background
- A de novo heterozygous c.320A>G [p.(His 107 Arg)] mutation in TUBA1A was identified in a patient with microcephaly, epileptic seizures, and severe developmental delay. PMID: 29109381
- Given that Spastin engages the MT in two places, we propose that severing occurs by forces exerted on the C-terminal tail of tubulin, which results in a conformational change in tubulin, leading to its release from the polymer. PMID: 17389232
- Molecular docking studies revealed that 6f interacted and bound efficiently with the colchicine-binding site of tubulin. Furthermore, 6f treatment induced G2/M cell cycle arrest in a dose-dependent manner, subsequently leading to cell apoptosis. PMID: 28440465
- Induced pluripotent stem cells (iPSCs) were generated from the umbilical cord and peripheral blood of two lissencephaly patients with varying clinical severities carrying alpha tubulin (TUBA1A) missense mutations. PMID: 27431206
- Long intergenic non-coding RNA APOC1P1-3 inhibits apoptosis by decreasing alpha-tubulin acetylation in breast cancer. PMID: 27228351
- Findings indicate that Tuba1a plays a crucial, non-compensated role in neuronal saltatory migration in vivo, highlighting the importance of microtubule flexibility in nucleus-centrosome coupling and neuronal-branching regulation during neuronal migration. PMID: 28687665
- Data suggest that TUBA1A mutations disrupting lateral interactions have pronounced dominant-negative effects on microtubule dynamics, which are associated with the severe end of the lissencephaly spectrum. PMID: 26493046
- Data demonstrate that tubulin phosphorylation and acetylation play significant roles in the regulation of microtubule assembly and stability. PMID: 26165356
- Data reveal that plasma membrane Ca(2+)-ATPase (PMCA) was associated with tubulin in both normotensive and hypertensive erythrocytes. PMID: 26307527
- Studies indicate that alpha-tubulin acetylation and microtubule levels are primarily governed by opposing actions of alpha-tubulin acetyltransferase 1 (ATAT1) and histone deacetylase 6 (HDAC6). PMID: 26227334
- Data from studies using a peptide fragment of alpha-tubulin (residues 31-49) suggest that Ser38 is essential for substrate recognition by alpha-tubulin acetylase 1 (ATAT1); Asp39, Ile42, the glycine stretch (residues 43-45), and Asp46 are also involved. PMID: 25602620
- Lysine 40 acetylation of alpha-tubulin does not result in significant changes in kinesin-1's landing rate or motility parameters. PMID: 24940781
- These results demonstrate that SelP interacts with tubulin, alpha 1a (TUBA1A). PMID: 24914767
- This study shows that all fetuses with lissencephaly and cerebellar hypoplasia carried distinct TUBA1A mutations. PMID: 25059107
- These findings highlight PKC-mediated phosphorylation of alpha-tubulin as a novel mechanism for controlling the dynamics of microtubules, ultimately affecting cell movement. PMID: 24574051
- This case provides new insights into the wide spectrum of disease phenotypes associated with TUBA1A mutations. PMID: 23528852
- The present study confirms that mutations in tubulin genes are responsible for complex brain malformation. PMID: 24392928
- Studies suggest that tubulin-interactive agents have the potential to play a significant role in the fight against cancer. PMID: 23818224
- Missense mutations in TUBA1A were found in 3 patients with polymicrogyria. PMID: 22948023
- We have described the clinical course and pathological findings in a child with a TUBA1A mutation. PMID: 22633752
- TUBA1A and TUBB2B coding regions have been sequenced, which are associated with cortical malformations. PMID: 23361065
- Data show that Na(+),K(+)-ATPase activity was greater than 50% lower, and membrane-associated tubulin content was greater than 200% higher in erythrocyte membranes from diabetic patients. PMID: 22565168
- This study describes a 14-month-old girl with TUBA1A mutation-associated lissencephaly and summarizes the clinical and neuroradiologic findings of 19 cases in the literature. PMID: 22264709
- Alpha2B-adrenergic receptor interaction with tubulin controls its transport from the endoplasmic reticulum to the cell surface. PMID: 21357695
- The expression of alpha-tubulin and MDR1 may play a significant role in the development and progression of human non-small cell lung carcinoma. PMID: 20510079
- We report a mutation in TUBA1A as a cause of polymicrogyria. While all mutations in TUBA1A have occurred de novo, resulting in isolated cases, this article describes familial recurrence of TUBA1A mutations due to somatic mosaicism in a parent. PMID: 21403111
- Data show that IAV-infected cells contain elevated levels of AcTub and alpha-tubulin. PMID: 21094644
- Mutations in TUBA1A result in defects in tubulin folding and heterodimer assembly. PMID: 20603323
- LIS-associated mutations of TUBA1A operate through diverse mechanisms that include disruption of binding sites for microtubule-associated proteins. PMID: 20466733
- The dipole moments of each tubulin isotype may influence their functional characteristics within the cell, leading to differences in MT assembly kinetics and stability. PMID: 16941085
- Mutations in alpha-tubulin in mice and humans that affect neuronal migration result in abnormal lamination of brain structures with associated behavioral deficits. PMID: 17218254
- Retrospective examination of MR images suggests that patients with TUBA1A mutations share not only cortical dysgenesis, but also cerebellar, hippocampal, corpus callosum, and brainstem abnormalities. PMID: 17584854
- Increased expression of tubulin alpha is associated with pulmonary sclerosing hemangioma. PMID: 17914564
- The diminished production of TUBA1A tubulin in R264C individuals is consistent with haploinsufficiency as a cause of the disease phenotype. PMID: 18199681
- The TUBA1A phenotype is distinct from LIS1, DCX, RELN, and ARX lissencephalies. Compared with the phenotypes of children mutated for TUBA1A, these prenatally diagnosed fetal cases occur at the severe end of the TUBA1A lissencephaly spectrum. PMID: 18669490
- Missense mutations within the TUBA1A gene are associated with specific abnormalities in lissencephaly. PMID: 18728072
- Mutation analysis in the TUBA1A gene was conducted in 46 patients with classical lissencephaly. PMID: 18954413
- This protein has been found differentially expressed in the Wernicke's Area from patients with schizophrenia. PMID: 19405953
Q&A
What criteria should guide selection of TUBA1A antibodies for basic cellular biology studies?
When selecting TUBA1A antibodies, researchers must prioritize:
-
Host species compatibility: Rabbit polyclonal antibodies (e.g., Boster Bio A03989 ) offer broad epitope recognition, while mouse monoclonal antibodies (e.g., Assay Genie MACO0009 ) provide higher specificity.
-
Application-specific validation: Antibodies validated for Western blot (WB) require distinct epitope accessibility compared to immunohistochemistry (IHC) or immunofluorescence (IF). For example, Boster Bio MA1107 demonstrates linear epitope recognition in WB but requires antigen retrieval for IHC .
-
Species reactivity: Confirm cross-reactivity using databases and manufacturer data. OriGene TA307921 reacts with bovine, chimpanzee, and human samples but shows limited utility in zebrafish .
Table 1: Key Selection Criteria for Common Applications
How should researchers validate TUBA1A antibody specificity in novel experimental systems?
A three-step validation protocol is recommended:
-
Knockdown/knockout controls: Compare signal intensity in TUBA1A-deficient cell lines versus wild-type.
-
Cross-reactivity profiling: Test against recombinant tubulin isoforms (e.g., TUBB2B, TUBA1B) using peptide blocking assays. Boster Bio antibodies show ≤5% cross-reactivity with non-target isoforms .
-
Subcellular localization: Confirm expected microtubule patterning via confocal microscopy. Discrepancies may indicate off-target binding, as observed in mutated TUBA1A models where antibodies failed to recognize cytoplasmic aggregates .
What are the methodological implications of TUBA1A’s post-translational modifications (PTMs) for antibody-based assays?
TUBA1A undergoes detyrosination, polyglutamylation, and acetylation, which impact antibody recognition:
-
Detyrosinated forms: Most commercial antibodies (e.g., OriGene TA307921) target the C-terminal epitope (aa 227–440 ), losing affinity in detyrosinated states.
-
Acetylated TUBA1A: Use antibodies specifically validated for modified forms, as standard antibodies may show reduced binding.
-
Quantification bias: PTM-rich samples (e.g., neuronal lysates) require normalization against total TUBA1A using antibodies against non-modifiable regions .
How can TUBA1A antibodies resolve contradictory data in mutation-associated microtubule instability studies?
The PMC study demonstrates a framework for analyzing TUBA1A mutants (e.g., R402C, C25F):
Methodological Workflow:
-
Transfection models: Express FLAG-tagged wild-type/mutant TUBA1A in HEK293 cells.
-
Dual-label IF: Combine anti-FLAG (1:1000) and anti-α-tubulin (1:5000 ) to differentiate incorporated vs. aggregated TUBA1A.
-
Microtubule density quantification:
Mutants showed 40–60% reduced density versus wild-type .
Table 2: Mutant-Specific Antibody Performance
| Mutation | Microtubule Incorporation (% Wild-Type) | Punctate Aggregates Observed |
|---|---|---|
| R64W | 72% | Rare |
| C25F | 58% | Frequent |
| R402C | 41% | Ubiquitous |
What advanced multiplexing strategies enhance TUBA1A dynamics studies in live-cell imaging?
Combine TUBA1A antibodies with:
-
Fluorescent biosensors: GFP-EB1 for microtubule plus-end tracking.
-
Photoactivatable tags: Perform fluorescence recovery after photobleaching (FRAP) to quantify turnover rates.
-
Critical controls:
-
Pre-absorb antibody with immunogen peptide (10x molar excess, 1 hr incubation) to confirm signal specificity.
-
Use lattice light-sheet microscopy to minimize phototoxicity during time-lapse imaging.
-
How should researchers interpret unexpected TUBA1A antibody reactivity in disease models?
Case example from neurodevelopmental disorders :
-
Observation: Anti-TUBA1A IHC showed reduced signal in lissencephaly patient brains despite normal mRNA levels.
-
Resolution workflow:
-
Alternate antibody validation: Compare with C-terminal vs. N-terminal antibodies.
-
Post-translational modification profiling: Mass spectrometry identified hyperdetyrosination (≥80% modified vs. 25% in controls).
-
Structural modeling: Mutation-induced epitope occlusion explained antibody binding failure.
-
What experimental parameters most significantly impact TUBA1A antibody performance in IHC?
A multivariate analysis of Boster Bio MA1107 data reveals:
Critical Factors (Ranked by Effect Size):
-
Antigen retrieval buffer pH: Citrate (pH 6.0) outperforms Tris-EDTA (pH 9.0) by 2.3-fold signal intensity.
-
Fixation time: Over-fixation (>48 hr in formalin) reduces epitope accessibility by 47%.
-
Blocking agent: 5% normal goat serum reduces non-specific binding by 81% versus BSA-only blocks.
How can researchers standardize TUBA1A quantification across WB platforms?
Implement a dual-normalization strategy:
-
Housekeeping protein: Use β-III tubulin (1:2000 dilution ) rather than GAPDH due to co-regulation in cytoskeletal studies.
-
Total protein normalization: Stain parallel gels with Coomassie Blue R-250 and quantify lane totals.
-
Cross-validated antibodies: Compare results from monoclonal (e.g., DM1A clone ) and polyclonal antibodies to control for lot-to-lot variability.
