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TUBA1A / Tubulin alpha-1A chain · Western blot design guide
Design a Western Blot for TUBA1A
Real validated TUBA1A Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TUBA1A WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
TUBA1A Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~50.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Skeletal muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated TUBA1A Western Blot Protocols
The A03989 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03989; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected TUBA1A Western Blot Band Size?
TUBA1A has a predicted mass of 50.1 kDa; isoforms and polyglutamylation could affect migration, but no distinct band pattern is demonstrated.
| Band near 50.1 kDa | consistent with the predicted TUBA1A mass; confirm identity with an independent antibody |
| Additional band at a different position | isoforms 1 and 2 could differ in migration, but resolution is unestablished |
| Band above 50.1 kDa | polyglutamylation at residues 443 and 445 could affect migration; its visible effect is unestablished |
| Weak band in soluble lysate | TUBA1A is a cytoskeletal protein that may remain in an insoluble fraction |
💡Expected TUBA1A appearanceUniProt predicts 50.1 kDa for TUBA1A; no empirical band size is supplied, and isoforms or modifications do not establish a specific migration shift, so confirm any candidate band with identity controls.
| UniProt predicted mass | places the reference size at 50.1 kDa |
| Isoform 1 | may migrate differently from isoform 2; its size is unspecified |
| Isoform 2 | may migrate differently from isoform 1; its size is unspecified |
| Polyglutamylation at residues 443 and 445 | could affect apparent migration; a visible shift is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytoskeletal TUBA1A may be poorly recovered from soluble lysate | examine the insoluble cytoskeletal fraction and verify extraction |
| Band higher than expected | polyglutamylation could affect migration, but the cause is unestablished | confirm TUBA1A identity with an independent antibody |
| Band lower than expected | isoform migration or protein fragmentation is possible but unestablished | check identity with a second antibody recognizing a different region |
| Multiple bands | isoforms 1 and 2 or modified forms could contribute; separation is unestablished | compare bands with an independent TUBA1A antibody |
| Weak or no signal | TUBA1A may remain in the insoluble cytoskeletal fraction | check fraction recovery and loading |
| Fragments below expected size | protein degradation during preparation is possible | prepare fresh lysate with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing TUBA1A band
A03989 · TUBA1A
A03989-Biotin · TUBA1A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TUBA1A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TUBA1A in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Skeletal muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Whole-tissue lysate controls are feasible because TUBA1A is cytoskeletal and HPA lists a not-detected tissue.
HPA tissue expression evidence for TUBA1A
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Medium | Protein (IHC) | HPA → |
Section 3
Advanced TUBA1A Western Blot Tips
Deeper troubleshooting and optimisation questions for TUBA1A, answered from its protein features.
How should TUBA1A band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could TUBA1A isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks canonical residues 1–35, so it may migrate differently from isoform 1. Check whether the antibody epitope includes that segment before interpreting a missing or additional band.
Which documented modifications could affect TUBA1A detection?
PTM · The supplied canonical coordinates list acetyllysine 40, nitrotyrosines 282 and 451, phosphoserine 439, and polyglutamylation at 443 and 445. Choose a modification-specific antibody only for the site being tested, and compare with total TUBA1A. These features alone do not establish a detectable band shift.
Report sites using the supplied UniProt canonical coordinates: 40, 282, 439, 443, 445 and 451. Isoform 2 lacks canonical residues 1–35, so its sequence numbering may differ. State the numbering convention when comparing antibody labels or publications.
Report sites using the supplied UniProt canonical coordinates: 40, 282, 439, 443, 445 and 451. Isoform 2 lacks canonical residues 1–35, so its sequence numbering may differ. State the numbering convention when comparing antibody labels or publications.
Does this guide establish induction of TUBA1A?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for TUBA1A?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03989 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should TUBA1A Western blot signals be quantified?
Quantitation · For total TUBA1A, quantify the band recognized by a total-protein antibody across consistently prepared samples. For a modification-specific signal, compare it with total TUBA1A from matched samples; changes in modification signal alone do not establish changes in total protein abundance.
Where should TUBA1A appear relative to its predicted mass?
Interpretation · The canonical TUBA1A sequence has a predicted mass of 50.1 kDa. No observed Western blot band is supplied, so use 50.1 kDa as a reference, not an expected apparent mass. The listed modifications do not establish a visible shift.
TUBA1A is annotated in the cytoplasmic cytoskeleton and flagellum axoneme. Keep sample preparation and fraction selection consistent across lanes, and compare like fractions when measuring its abundance.
Check antibody epitope coverage against isoform 2’s missing canonical residues 1–35 and consider whether the antibody recognizes a listed modification. TUBA1A also forms alpha–beta tubulin dimers. The supplied features cannot identify an unexpected band or prove that a modification caused its mobility.
TUBA1A is annotated in the cytoplasmic cytoskeleton and flagellum axoneme. Keep sample preparation and fraction selection consistent across lanes, and compare like fractions when measuring its abundance.
Check antibody epitope coverage against isoform 2’s missing canonical residues 1–35 and consider whether the antibody recognizes a listed modification. TUBA1A also forms alpha–beta tubulin dimers. The supplied features cannot identify an unexpected band or prove that a modification caused its mobility.
Boster reagents
TUBA1A Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-alpha-Tubulin TUBA1A Antibody
Cat # A03989
Real WB data
Anti-Alpha-tubulin TUBA1A Antibody (biotin)
Cat # A03989-Biotin
The catalog reports two anti-TUBA1A antibodies with Western blot images: A03989 and biotinylated A03989-Biotin. Their captions show cell and tissue lysates tested at 1 μg/mL. The pictured specimens document those specific contexts, not universal validation across species.
Which to pick: Choose A03989 for listed human, mouse, or rat reactivity. Choose A03989-Biotin if you need a biotinylated antibody or listed chicken, rabbit, or zebrafish reactivity. Both have Western blot images; check each caption for the samples tested.
Source: BosterBio TUBA1A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q71U36.
- Human Protein Atlas. TUBA1A tissue expression.
- PMC4929438 — target-verified WB comparison
- PMC12687654 — target-verified WB comparison
- PMC10332271 — target-verified WB comparison