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- Table of Contents
SMAD7 · Western blot design guide
SMAD7 Western Blot Planning Guide
Plan a SMAD7 Western blot around the catalog-observed 46.4 kDa band, image-backed A00784-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SMAD7 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 | 46.4 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Adipose tissue | |
| Negative control | Parathyroid gland |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 46.4 kDa |
| ⓘ | Localization | Nucleus / Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated SMAD7 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adipose tissue |
| 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 | A00784-1 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 SMAD7 Western Blot Band Size?
Use the product-observed 46.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 46.4 kDa | Matches the authoritative product WB observation. |
| 46.4 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 SMAD7 appearancePlan around 46.4 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 46.4 kDa; use this as the primary experimental expectation. |
| Calculated mass | 46.4 kDa from UniProt O15105; 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 A00784-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 46.4 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 SMAD7 band
A00784-1 · SMAD7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SMAD7 Western blot
🧪Use Adipose tissue as the first positive-control candidate and Parathyroid gland as the HPA Not detected negative candidate.
Positive control: Adipose tissue (Medium)
Negative control: Parathyroid gland (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for SMAD7
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 |
|---|---|---|---|---|
| Adipose tissue | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Caudate | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Parathyroid gland | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
Section 3
Advanced SMAD7 Western Blot Tips
Deeper troubleshooting and optimisation questions for SMAD7, answered from its protein features.
Which band should guide the blot?
Use 46.4 kDa, the observation attached to the authoritative A00784-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 46.4 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Parathyroid gland 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 SMAD7 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00784-1 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
SMAD7 Western Blot Reagents
Human/Mouse/Rat-reactive SMAD7 Western blot reagents with authoritative product imagery.
Real WB data
Anti-MADH7/SMAD7 Antibody Picoband®
Cat # A00784-1
Real WB data
Anti-MADH7/SMAD7 Antibody Picoband®
Cat # A00784-2
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 O15105
- Human Protein Atlas — SMAD7
- A00784-1 product record
- PMC6522533 — Smad7:β-catenin complex regulates myogenic gene transcription (Cell death & disease, 2019)
- PMC12573700 — [Expression and prognostic value of mothers against decapentaplegic homolog 7 in head and neck squamous cell carcinoma] (Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology, 2025)
- PMC3220473 — The deubiquitinase CYLD targets Smad7 protein to regulate transforming growth factor β (TGF-β) signaling and the development of regulatory T cells (The Journal of biological chemistry, 2011)