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- Table of Contents
TUB · Western blot design guide
TUB Western Blot Planning Guide
Plan a TUB Western blot around the catalog-observed 55.7 kDa band, image-backed A02917-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
TUB Western Blot Experimental Design Guide
Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | 55.7 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Cerebral cortex | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 55.7 kDa |
| ⓘ | Localization | Cytoplasm / Nucleus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated TUB Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Cerebral cortex |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A02917-1 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected TUB Western Blot Band Size?
Use the product-observed 55.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 55.7 kDa | Matches the authoritative product WB observation. |
| 55.7 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected TUB appearancePlan around 55.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 55.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 55.7 kDa from UniProt P50607; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A02917-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 55.7 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing TUB band
A02917-1 · TUB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TUB Western blot
🧪Use Cerebral cortex as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.
Positive control: Cerebral cortex (High)
Negative control: Bone marrow (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for TUB
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebral cortex | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cervix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced TUB Western Blot Tips
Deeper troubleshooting and optimisation questions for TUB, answered from its protein features.
Which band should guide the blot?
Use 55.7 kDa, the observation attached to the authoritative A02917-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 55.7 kDa expectation.
Which positive control should I start with?
Start with Cerebral cortex, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Bone marrow as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for TUB Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02917-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents
TUB Western Blot Reagents
Human-reactive TUB Western blot reagents with authoritative product imagery.
Real WB data
Anti-TUB 1 Antibody Picoband®
Cat # A02917-1
Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.
Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.
References
- UniProt P50607
- Human Protein Atlas — TUB
- A02917-1 product record
- PMC4091484 — Insulin Phosphorylates Tyrosine Residue 464 of Tub and Translocates Tubby into the Nucleus in HIRcB Cells (Endocrinology and metabolism (Seoul, Korea), 2014)