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- Table of Contents
TUBB2A / Tubulin beta-2A chain · Western blot design guide
Design a Western Blot for TUBB2A
Source-linked TUBB2A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TUBB2A 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
TUBB2A Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~49.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Placenta (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked TUBB2A Western Blot Protocol Options
The M06868 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 | M06868; 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 TUBB2A Western Blot Band Size?
TUBB2A is predicted at 49.9 kDa; listed modifications could affect migration, but no empirical band size or visible shift is demonstrated.
| Band near 49.9 kDa | Consistent with the predicted full-length TUBB2A size; confirm identity with antibody controls |
| Closely spaced doublet near 50 kDa | Modified TUBB2A is possible, but distinct migration is not established |
| Band slightly above 50 kDa | Polyglutamylation is possible, but its effect on migration is not established |
| Weak band in a soluble fraction | Cytoskeletal TUBB2A may be underrepresented in that fraction |
💡Expected TUBB2A appearanceUniProt predicts a 49.9 kDa TUBB2A band; no empirical band size or demonstrated migration shift is supplied, so confirm any candidate band with antibody specificity controls.
| Predicted molecular mass | Places full-length TUBB2A near 49.9 kDa |
| Phosphorylation at Ser40 and Ser172 | Adds modification mass, but a visible band shift is not established |
| Alternate acetylation or succinylation at Lys58 | Changes mass slightly; distinct migration is not established |
| Polyglutamylation at residue 438 | Can add variable mass; its effect on this blot is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeletal TUBB2A may be poorly recovered | Check the insoluble fraction and confirm lysis efficiency |
| Band higher than expected | A modified species is possible, but its identity is unproven | Compare with a validated TUBB2A antibody and an appropriate modification control |
| Band lower than expected | Protein breakdown or a nonspecific band is possible | Use protease inhibitors and confirm band identity with an independent antibody |
| Multiple bands | Modified species or antibody cross-reactivity may contribute | Check antibody specificity and compare treated and untreated samples |
| Weak or no signal | Cytoskeletal protein recovery or antibody detection may be insufficient | Check fraction recovery, loading, and a positive lysate control |
Experiment evidence
Western blot images showing TUBB2A band
M06868 · TUBB2A
M06868 · TUBB2A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TUBB2A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TUBB2A in Western blot, you can use placenta tissue lysate.
Positive control: Placenta (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Its cytoskeletal location supports tissue lysate testing, and HPA lists adipose tissue as not detected.
HPA tissue expression evidence for TUBB2A
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 |
|---|---|---|---|---|
| Placenta | endothelial cells | High | Protein (IHC) | HPA → |
| Skin | cells in granular layer | High | Protein (IHC) | HPA → |
| Testis | pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TUBB2A Western Blot Tips
Deeper troubleshooting and optimisation questions for TUBB2A, answered from its protein features.
Where should the TUBB2A band appear relative to predicted mass?
Band shift · The predicted mass is 49.9 kDa. No observed band position is supplied, so compare your signal with a molecular weight marker rather than assuming an apparent mass. The listed modifications alone do not establish a visible shift.
Could a second TUBB2A band represent a listed isoform?
Isoforms · Only one isoform is listed, with no alternative sequence. A second band cannot be assigned to a TUBB2A isoform from these features alone.
Which TUBB2A modifications matter when interpreting band shifts?
PTM · Listed sites include phosphoserine 40 and 172, phosphothreonine 285 and 290, alternate acetylation or succinylation at lysine 58, methylarginine 318, and 5-glutamyl polyglutamate at 438. These are UniProt coordinates. Their presence does not establish a visible shift or explain a measured mass difference.
UniProt lists phosphoserine 172 as modified by CDK1. A phospho-specific assay targeting that site may reflect its phosphorylation state; a total-TUBB2A blot cannot establish that state. Serine 172 uses the supplied UniProt numbering.
UniProt lists phosphoserine 172 as modified by CDK1. A phospho-specific assay targeting that site may reflect its phosphorylation state; a total-TUBB2A blot cannot establish that state. Serine 172 uses the supplied UniProt numbering.
Does this guide establish induction of TUBB2A?
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.
What transfer method to use for TUBB2A Western blot?
Transfer · TUBB2A has a predicted mass of 49.9 kDa. Choose transfer conditions suitable for a protein near 50 kDa and verify transfer at that size. The supplied features do not specify a membrane or transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M06868 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 TUBB2A signals be quantified across samples?
Quantitation · Measure the TUBB2A signal in the linear detection range and normalize consistently across samples. If comparing a site-specific modification, report its signal relative to total TUBB2A; the listed modification sites do not establish how much protein is modified.
What could explain unexpected bands in a TUBB2A blot?
Interpretation · Check whether each band is near the predicted 49.9 kDa position and confirm antibody specificity. UniProt lists one isoform and several modification sites, including polyglutamylation at 438, but those features alone cannot identify an unexpected band or establish a visible shift.
Boster reagents
TUBB2A 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-beta II tubulin TUBB2A Monoclonal Antibody
Cat # M06868
M06868 is a monoclonal anti-TUBB2A antibody listed for human, mouse, and rat. Its Western blot image shows mouse and rat brain tissue at 1:100000 dilution; no human sample is shown in the supplied evidence.
Which to pick: M06868 is the only listed option. Its Western blot image provides a practical starting point for mouse or rat brain tissue; human reactivity is listed, but the supplied image does not show a human sample.
Source: BosterBio TUBB2A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13885.
- Human Protein Atlas. TUBB2A tissue expression.
- PMC4502645 — target-verified WB comparison
- PMC7537480 — target-verified WB comparison
- PMC4873365 — target-verified WB comparison