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- Table of Contents
TEAD4 / Transcriptional enhancer factor TEF-3 · Western blot design guide
Design a Western Blot for TEAD4
Real validated TEAD4 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TEAD4 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
TEAD4 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 | ~48.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated TEAD4 Western Blot Protocols
The A04289-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04289-1) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04289-1) |
| Primary antibody | A04289-1 · 1:1000 (catalog A04289-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04289-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04289-1) |
Section 2
What Is the Expected TEAD4 Western Blot Band Size?
TEAD4 is predicted at 48.3 kDa; three isoforms could affect migration, but no empirical band or distinct isoform pattern is supplied.
| Band near 48.3 kDa | Consistent with the predicted TEAD4 mass; confirm identity with controls |
| Several bands near the expected region | TEAD4 has three named isoforms, but their migration is unknown |
| One band despite three isoforms | The isoforms are not known to resolve into distinct bands |
| Weak or absent band in a cytoplasmic fraction | TEAD4 is annotated as nuclear |
💡Expected TEAD4 appearanceTEAD4 has a predicted mass of 48.3 kDa, but no empirical band size is supplied; use an appropriate positive control and TEAD4 depletion to assess band identity.
| Predicted TEAD4 mass | Sets a reference of 48.3 kDa, not a validated migration position |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Alternative splicing may affect size, but its individual mass and migration are not supplied |
| Isoform 3 | Alternative splicing may affect size, but its individual mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TEAD4 signal may be below detection in the sampled lysate | Check a positive control and compare whole-cell with nuclear extract |
| Weak or no signal | Nuclear TEAD4 may be underrepresented in the sampled fraction | Check nuclear extraction and load a positive control |
| Multiple bands | TEAD4 has three isoforms, though distinct migration is unconfirmed | Test which bands decrease after TEAD4 depletion |
| Band higher than expected | Isoform identity or band identity is unresolved | Compare with a positive control and test TEAD4 depletion |
| Band lower than expected | Isoform identity or band identity is unresolved | Compare with a positive control and test TEAD4 depletion |
Experiment evidence
Western blot images showing TEAD4 band
A04289-1 · TEAD4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TEAD4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TEAD4 in Western blot, you can use appendix tissue, which HPA rates as medium for TEAD4.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: TEAD4 is nuclear, so nuclear-enriched lysate may improve detection; HPA identifies adipose tissue as not detected.
HPA tissue expression evidence for TEAD4
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 |
|---|---|---|---|---|
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TEAD4 Western Blot Tips
Deeper troubleshooting and optimisation questions for TEAD4, answered from its protein features.
How should TEAD4 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 TEAD4 isoforms produce different bands?
Isoforms · Three isoforms are listed. In UniProt coordinates, isoform 2 lacks residues 1–129 and isoform 3 lacks residues 119–161. These sequence differences could affect band size, but the features do not establish where either isoform migrates.
Check the antibody’s binding region against the listed deletions. An antibody requiring UniProt residues 1–129 may miss isoform 2; one requiring residues 119–161 may miss isoform 3. Confirm the antibody’s numbering convention before comparing coordinates.
Check the antibody’s binding region against the listed deletions. An antibody requiring UniProt residues 1–129 may miss isoform 2; one requiring residues 119–161 may miss isoform 3. Confirm the antibody’s numbering convention before comparing coordinates.
Can an annotated modification explain a TEAD4 band shift?
PTM · This record lists no modified residues or glycosylation sites. Do not assign a shifted band to a specific modification from these features. A predicted mass alone also cannot establish the cause of a migration difference.
Does this guide establish induction of TEAD4?
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 TEAD4?
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 A04289-1 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 TEAD4 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
What size band should I expect for TEAD4?
Interpretation · The canonical TEAD4 sequence is 434 residues with a predicted mass of 48.3 kDa. No observed band position is supplied, so use 48.3 kDa as a reference, not a guaranteed migration position.
Compare its position with the 48.3 kDa canonical prediction, then check whether the antibody can detect isoforms 2 and 3. The listed deletions make isoforms worth considering, but these features alone cannot identify an unexpected band.
TEAD4 is listed as nuclear and has three isoforms. Use a consistent sample preparation and decide whether the measurement concerns one band or the combined TEAD4 signal. Keep that choice consistent across samples.
Compare its position with the 48.3 kDa canonical prediction, then check whether the antibody can detect isoforms 2 and 3. The listed deletions make isoforms worth considering, but these features alone cannot identify an unexpected band.
TEAD4 is listed as nuclear and has three isoforms. Use a consistent sample preparation and decide whether the measurement concerns one band or the combined TEAD4 signal. Keep that choice consistent across samples.
Boster reagents
TEAD4 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-TEAD4 Antibody
Cat # A04289-1
The catalog reports one anti-TEAD4 antibody, A04289-1, with reported Human, Mouse, and Rat reactivity. Its Western blot image uses extracts from various cell lines at 1:1000 and 25 µg per lane; the caption does not identify the cell lines.
Which to pick: A04289-1 is the only listed option and has a Western blot image. Check its reported reactivity against your sample species; the image caption does not establish which species were tested.
Source: BosterBio TEAD4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15561.
- Human Protein Atlas. TEAD4 tissue expression.
- PMC12518352 — target-verified WB comparison
- PMC9112458 — target-verified WB comparison
- PMC5623908 — target-verified WB comparison