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- Table of Contents
TNFSF12 · Western blot design guide
TNFSF12 Western Blot Planning Guide
Plan a TNFSF12 Western blot around the catalog-observed 27.2 kDa band, image-backed A02009-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
TNFSF12 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 | 27.2 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Adrenal gland | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 27.2 kDa |
| ⓘ | Localization | Cell membrane / Secreted |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated TNFSF12 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A02009-1 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 TNFSF12 Western Blot Band Size?
Use the product-observed 27.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 27.2 kDa | Matches the authoritative product WB observation. |
| 27.2 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 TNFSF12 appearancePlan around 27.2 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 27.2 kDa; use this as the primary experimental expectation. |
| Calculated mass | 27.2 kDa from UniProt O43508; 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 A02009-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 27.2 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 TNFSF12 band
A02009-1 · TNFSF12
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TNFSF12 Western blot
🧪Use Adrenal gland as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.
Positive control: Adrenal gland (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 TNFSF12
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 |
|---|---|---|---|---|
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Duodenum | Reported tissue cells | High | Protein (HPA) | HPA → |
| Heart muscle | Reported tissue cells | High | Protein (HPA) | HPA → |
| Rectum | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced TNFSF12 Western Blot Tips
Deeper troubleshooting and optimisation questions for TNFSF12, answered from its protein features.
Which band should guide the blot?
Use 27.2 kDa, the observation attached to the authoritative A02009-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 27.2 kDa expectation.
Which positive control should I start with?
Start with Adrenal gland, 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 TNFSF12 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02009-1 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
TNFSF12 Western Blot Reagents
Human/Mouse/Rat-reactive TNFSF12 Western blot reagents with authoritative product imagery.
Real WB data
Anti-TWEAK/TNFSF12 Antibody Picoband®
Cat # A02009-1
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 O43508
- Human Protein Atlas — TNFSF12
- A02009-1 product record
- PMC11794540 — Endothelial AGGF1 promotes retinal angiogenesis by coordinating TNFSF12/FN14 signalling (Nature communications, 2025)
- PMC12477863 — Expression and significance of TNFSF12/TWEAK, CD163 in monocytes of patients with newly diagnosed rheumatoid arthritis (Biomarkers in medicine, 2025)
