The antibody is provided as a liquid solution in phosphate-buffered saline (PBS) containing 50% glycerol, 0.5% bovine serum albumin (BSA), and 0.02% sodium azide.
Form
Liquid
Lead Time
We typically dispatch orders within 1-3 business days of receipt. Delivery timelines may vary depending on the purchasing method and location. For specific delivery estimates, please consult your local distributor.
What are the key differences between the major alpha-tubulin isoforms targeted by antibodies?
Alpha-tubulin isoforms have distinct expression patterns and functions despite high sequence homology:
Isoform
Primary Expression
Key Function
Associated Conditions
Molecular Weight
TUBA1A
Central nervous system, highest in developing brain neurons
Neuronal migration, brain development
Lissencephaly, neurodevelopmental disorders
50-55 kDa
TUBA1B
Ubiquitous, particularly in proliferating cells
Microtubule stability, cell division
Cancer biomarker, prognostic factor
50-55 kDa
TUBA1C
Broadly expressed, upregulated in various cancers
Cell proliferation, cell cycle progression
Pancreatic cancer, glioblastoma
50-55 kDa
TUBA3C
Widely expressed
Structural component of microtubules
Less characterized
50-55 kDa
TUBA4A
Widespread, enriched in specific neuronal populations
Microtubule assembly and stability
Amyotrophic Lateral Sclerosis (ALS)
55 kDa
TUBA1A is highly expressed in post-mitotic neurons in the developing brain and 100% identical to its rat homolog, differing by only 2-3 amino acids from pig and chicken homologs.
What are the optimal conditions for using alpha-tubulin antibodies in Western blotting?
When using alpha-tubulin antibodies for Western blotting, researchers should consider:
Parameter
Recommendation
Notes
Sample preparation
Resuspend ~50 million cells in 1 mL cold lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF with protease inhibitor cocktail)
Incubate 60 min on ice, then centrifuge to remove debris
Dilution range
1:3000-1:12000 for WB, optimal dilution is antibody-specific
Example: Anti-TUBA1B (11224-1-AP) works well at 1:3000-1:12000
Positive controls
Cell lines: HEK-293, HeLa, A549, NIH/3T3; Tissues: rat brain
Choose appropriate controls for your species of interest
Blocking solution
3% BSA in PBST
Recommended for reduced background
Incubation time
Primary antibody: overnight at 4°C; Secondary antibody: 1 hr at room temperature
Adjust based on signal strength
Expected MW
50-55 kDa
Single band should be visible at this range
For optimal results, "Some samples may show large bands due to abundant expression of tubulin in certain tissues and cell types. Adjustment of loading amount or contrast may be necessary for clear visualization".
How should alpha-tubulin antibodies be stored to maintain optimal activity?
Proper storage is critical for maintaining antibody performance:
Storage Condition
Recommended for
Notes
-20°C
Long-term storage (up to 1 year from receipt)
For lyophilized antibodies
4°C
After reconstitution (up to 1 month)
For frequent use
-20°C
After reconstitution (up to 6 months, aliquoted)
Avoid repeated freeze-thaw cycles
"The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition".
What considerations are important when using alpha-tubulin antibodies as loading controls?
Alpha-tubulin is commonly used as a loading control in Western blots:
"Like GAPDH and beta-Actin, alpha-tubulin antibodies make excellent loading controls in immunoblots"
Advantages over other controls: "The protein is better than GAPDH for quantification in heart diseases"
Recommended loading amount: 10-30 μg of total protein per lane
Consideration: "Even lower dilution can be used" when high sensitivity is not required
Important note: Expression levels may vary across tissue types, so validation in your specific experimental system is essential
How can researchers ensure antibody specificity when working with highly conserved alpha-tubulin isoforms?
Distinguishing between alpha-tubulin isoforms requires careful consideration:
Approach
Methodology
Considerations
Epitope selection
Choose antibodies targeting unique regions
The N-terminal structural domain (aa 65-97) is recognized by some pan-alpha-tubulin antibodies
Cross-reactivity testing
Validate against multiple species
Some antibodies like anti-TUBA1A DM1A clone react with chicken, human, mouse, and rat samples
Immunogen verification
Check immunogen used for antibody production
Example: "Microtubules from chicken embryo brain" for DM1A clone
BLAST analysis
Compare sequence homology
"Run a BLAST between your target species and the immunogen sequence to predict cross-reactivity"
Validation controls
Include positive and negative controls
Use samples with known expression patterns of specific isoforms
"The antibody recognizes the defined epitope (aa 65-97) on N-terminal structural domain of alpha-tubulin. Reacts with all species (recognized epitope conserved within all species)" - this highlights the challenge of isoform specificity due to conservation.
What methodologies are most effective for studying TUBA1A mutations associated with neurodevelopmental disorders?
TUBA1A mutations are linked to neurodevelopmental disorders including lissencephaly:
Methodology
Application
Research Insights
In utero electroporation
Expressing TUBA1A mutants in developing mouse brain
"Ectopic expression of TUBA1A-V409I/A mutants disrupt neuronal migration in mice and promote excessive neurite branching"
Microtubule assembly assays
Testing mutant incorporation into microtubules
"Some TUBA1A mutants appear to be gain-of-function, as they are capable of microtubule assembly and act dominantly to perturb microtubule function in migrating neurons"
Immunohistochemistry
Visualizing neuronal migration defects
TUBA1A antibodies can detect abnormal neuronal positioning in cortical samples
Genetic screening
Identifying novel mutations
"To date, a total of 121 heterozygous, missense mutations have been identified in TUBA1A"
Computational modeling
Predicting mutation effects
Structural analysis can help determine if mutations affect tubulin folding, dimer formation, or microtubule stability
"It remains unclear whether the different severities of malformations observed in these patients are a result of differences in genetic background or are a result of a specific functional difference in the mutant tubulin".
How can alpha-tubulin antibodies be used to investigate microtubule dynamics in cancer research?
Alpha-tubulin isoforms have emerged as important cancer biomarkers:
Research Application
Methodology
Key Findings
Prognostic biomarker identification
Immunohistochemistry on tumor microarrays
"TUBA1C was overexpressed in most cancers, and overexpression of TUBA1C was linked to poor prognosis and higher tumour grade in patients"
Immune microenvironment analysis
Single-sample GSEA (ssGSEA) + immunostaining
"TUBA1C expression was associated with tumour mutation burden (TMB), microsatellite instability (MSI), the tumour microenvironment (TME) and the infiltration of immune cells"
Cancer cell migration studies
Live cell imaging with fluorescently tagged antibodies
Tubulin dynamics correlate with metastatic potential in multiple cancer types
Drug sensitivity prediction
Correlation analysis of expression data
"The sensitivity of 10 anticancer drugs was associated with high TUBA1C expression"
Immune checkpoint therapy prediction
ROC curve analysis
"The group with high TUBA1B responded better to ICI therapy than the group with low TUBA1B in LGG, LIHC, LUAD and LUSC"
"In pancreatic cancer, TUBA1C knockdown significantly inhibited cell proliferation and metastasis, suggesting its potential as a therapeutic target in PDAC".
What are the methodological considerations when using TUBA4A antibodies to study Amyotrophic Lateral Sclerosis (ALS)?
TUBA4A mutations have been implicated in ALS pathogenesis:
Consideration
Methodology
Research Findings
Mutation detection
Molecular dynamic (MD) modeling
"Each of the reported mutations will cause notable structural changes to the TUBA4A (α chain) tertiary protein structure, adversely affecting its physical properties and functions"
GTP binding assessment
Molecular docking and MD simulations
"Certain α chain mutations (e.g. K430N, R215C, and W407X) may cause structural deviations that impair GTP binding, and plausibly prevent or destabilize tubulin polymerization"
Aggregation propensity
Computational prediction models
"Several mutations (including R320C and K430N) confer a significant increase in predicted aggregation propensity of TUBA4A mutants relative to wild-type"
Microtubule assembly
In vitro polymerization assays
TUBA4A mutations affect α-tubulin::β-tubulin dimerization, GTP binding, and microtubule assembly and stability
Post-translational modifications
Mass spectrometry
"TUBA4A comprises a folded core flanked by two intrinsically disordered tails that are 'hotspots' for post-translational modification"
"Studies to determine the mechanism of neurotoxicity and the impact of ALS-linked mutations in TUBA4A are critically needed for developing therapeutic interventions".
What immunological insights can be gained from studying alpha-tubulin expression in cancer?
Recent research has revealed important connections between alpha-tubulin isoforms and cancer immunity:
Research Focus
Methodology
Key Findings
Tumor Mutation Burden (TMB) correlation
Bioinformatic analysis of TCGA data
"A positive association between TUBA1C expression and TMB was observed in 15 cancers, including UCEC, STAD, SKCM, LUAD, LGG, and others"
Microsatellite Instability (MSI)
Computational analysis
"In ACC, UCEC, SARC and COAD, the TUBA1C expression level was positively associated with MSI"
Tumor Microenvironment (TME)
ESTIMATE algorithm
"TUBA1C expression was positively correlated with stromal scores in GBM and LGG, while a negative correlation was observed in ESCA, STAD and TGCT"
Immune checkpoint correlation
Expression correlation analysis
"In OV, LGG, BRCA and BLCA, a positive association of TUBA1C expression with immune checkpoints was revealed"
Immune cell infiltration
Timer2.0 database analysis
"TUBA1C expression was positively related to M1 macrophage infiltration in CESC, KIRP, LGG and UCEC"
"Our study indicates that TUBA1B could potentially serve as a diagnostic marker for predicting cancer immunological profiles and survival outcomes".
Why might researchers observe inconsistent results when using alpha-tubulin antibodies across different applications?
Several factors can affect the performance of alpha-tubulin antibodies:
Issue
Possible Cause
Solution
Weak or no signal in Western blot
Insufficient protein loading
"Some samples had big bands due to the large amounts present in the samples"; adjust loading accordingly
Multiple bands
Cross-reactivity with other tubulin isoforms
"Cross-reactivity with other proteins" can occur; validate antibody specificity
Variable signal across tissue types
Different expression levels
"TUBA1A is expressed in a wide range of tissues but shows higher levels in the central nervous system"
Poor signal in fixed tissues
Epitope masking
"For immunohistochemistry, suggested antigen retrieval with TE buffer pH 9.0; alternatively, antigen retrieval may be performed with citrate buffer pH 6.0"
Inconsistent results between species
Sequence variation
Researchers report: "Works well when diluted in 1X TBST" for better consistency
"The antibody works well for detection of Tubulin by Western blot at 0.5 and 1 μg/mL", but optimal conditions may vary based on the specific antibody and application.
How can researchers optimize immunofluorescence protocols when using alpha-tubulin antibodies?
Immunofluorescence is a common application for alpha-tubulin antibodies:
Parameter
Recommendation
Notes
Fixation method
4% paraformaldehyde
"Immunocytochemistry labelling of (4% PFA) fixed SH-SY5Y cells by Alpha Tubulin Polyclonal antibody at dilution of 1:100 showed strong labelling"
Dilution range
1:50-1:500 for IF/ICC
Dilution depends on specific antibody and sample type
Permeabilization
0.1-0.5% Triton X-100
Required for accessing intracellular tubulin structures
Blocking solution
3-5% BSA or normal serum
Reduces non-specific binding
Counterstaining
DAPI for nuclei
Helps with cellular localization
Validated cell lines
HeLa, HepG2, MCF-7, Lymphatic endothelial cells
"Very nice staining for A549 cells with 1:200 dilution"
What approaches can be used to validate the specificity of antibodies targeting specific alpha-tubulin isoforms?
Antibody validation is critical for ensuring reliable results:
Validation Approach
Methodology
Consideration
Knockout/knockdown controls
siRNA or CRISPR-Cas9
"The sequences of siRNA1 (TUBA1C-si1), siRNA2 (TUBA1C-si2), and siRNA3 (TUBA1C-si3) were as follows: siRNA1 5′-GAACCUGGCUGUGAUUCAATT-3′; siRNA2 5′-CGAACAGCUUACUGUAGCATT-3′; and siRNA3 5′-GGAAUUAGAUCCUUCAAAUTT-3′"
Peptide competition assay
Pre-incubate antibody with immunizing peptide
"Blocking peptide can be purchased. Costs vary based on immunogen length"
Multiple antibody comparison
Use antibodies targeting different epitopes
Compare results from antibodies recognizing N-terminal vs. C-terminal regions
"Our lab technicians have not tested anti-Alpha-Tubulin antibody (Monoclonal, DM1A) MA1107 on monkey. You can run a BLAST between monkey and the immunogen sequence of anti-Alpha-Tubulin antibody (Monoclonal, DM1A) MA1107 to see if they may cross-react".
How can alpha-tubulin antibodies be used to advance understanding of cancer biomarkers and therapeutic targets?
Alpha-tubulin isoforms show promise as cancer biomarkers:
Research Direction
Methodology
Findings
Diagnostic biomarker development
ROC curve analysis
"The area under the curve (AUC) serves as a diagnostic performance indicator, with values closer to 1 indicating higher diagnostic accuracy. AUC values between 0.7 and 0.9 denote moderate diagnostic capacity for TUBA1B"
Prognostic nomogram development
Cox regression analysis
"Factors with p < 0.1 in the univariate analysis were included in the multivariate Cox analysis" to build predictive models
Immunotherapy response prediction
Correlation with immune checkpoint expression
TUBA1C "was correlated with immune checkpoints in all cancers except UVM, KIRC and ACC"
Drug sensitivity screening
Pharmacogenomic analysis
Association between TUBA1C expression and sensitivity to anticancer drugs
Tumor microenvironment characterization
Single-sample GSEA (ssGSEA)
"ssGSEA algorithm was used to evaluate the infiltration levels of 24 immune cell types across pan-cancer"
"TUBA1C might be a biomarker for predicting the immune status and prognosis of cancers, offering new ideas for cancer treatment".
What are the future research directions for studying alpha-tubulin mutations in neurodevelopmental disorders?
Understanding alpha-tubulin's role in neurodevelopment remains an important research frontier:
Research Direction
Methodology
Significance
Isoform-specific functions
Conditional knockout models
Understanding why TUBA1A mutations specifically affect brain development
Mechanistic classification
In vitro and in vivo functional assays
"It is important to establish clear mechanistic themes that will be useful in predicting the molecular, cellular, and tissue-level phenotypes of specific mutations"
Therapeutic intervention
Small molecule screening
Identifying compounds that can rescue neurodevelopmental defects
Genotype-phenotype correlations
Patient cohort studies
"It remains unclear whether the different severities of malformations observed in these patients are a result of differences in genetic background or are a result of a specific functional difference in the mutant tubulin"
Post-translational modification analysis
Proteomic approaches
Understanding how modifications affect tubulin function in neurodevelopment
How can advanced imaging techniques using alpha-tubulin antibodies enhance our understanding of cellular processes?
Innovative imaging approaches with alpha-tubulin antibodies:
Imaging Technique
Application
Research Potential
Super-resolution microscopy
Visualizing microtubule dynamics
Resolving individual microtubules below the diffraction limit
Live-cell imaging
Tracking microtubule formation and disassembly
Understanding dynamic instability in real-time
Proximity ligation assay
Detecting protein-protein interactions
Identifying tubulin-interacting proteins
FRET-based sensors
Measuring conformational changes
Tracking post-translational modifications
Expansion microscopy
Enhanced visualization of cytoskeletal networks
Physical expansion of samples for improved resolution
These techniques, combined with specific alpha-tubulin isoform antibodies, can provide unprecedented insights into microtubule biology in health and disease.
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