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- Table of Contents
TRAF3 · Western blot design guide
TRAF3 Western Blot Planning Guide
Plan a TRAF3 Western blot around the catalog-observed 64.5 kDa band, image-backed A00424 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
TRAF3 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 | 64.5 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Small intestine | |
| Negative control | Breast |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 64.5 kDa |
| ⓘ | Localization | Cytoplasm / Endosome |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated TRAF3 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Pancreas |
| 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 | A00424 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 TRAF3 Western Blot Band Size?
Use the product-observed 64.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 64.5 kDa | Matches the authoritative product WB observation. |
| 64.5 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 TRAF3 appearancePlan around 64.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 64.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 64.5 kDa from UniProt Q13114; 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 A00424. |
| Situation | Likely cause | Next action |
|---|---|---|
| 64.5 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 TRAF3 band
A00424 · TRAF3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TRAF3 Western blot
🧪Use Small intestine as the first positive-control candidate and Breast as the HPA Not detected negative candidate.
Positive control: Small intestine (High)
Negative control: Breast (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for TRAF3
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 |
|---|---|---|---|---|
| Small intestine | Reported tissue cells | High | Protein (HPA) | HPA → |
| Pancreas | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Caudate | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced TRAF3 Western Blot Tips
Deeper troubleshooting and optimisation questions for TRAF3, answered from its protein features.
Which band should guide the blot?
Use 64.5 kDa, the observation attached to the authoritative A00424 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 64.5 kDa expectation.
Which positive control should I start with?
Start with Small intestine, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Breast 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 TRAF3 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00424 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
TRAF3 Western Blot Reagents
Human/Mouse/Rat-reactive TRAF3 Western blot reagents with authoritative product imagery.
Real WB data
Anti-TRAF3 Antibody Picoband®
Cat # A00424
Real WB data
Anti-TRAF3 Antibody Picoband®
Cat # PA2008
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 Q13114
- Human Protein Atlas — TRAF3
- A00424 product record
- PMC2766052 — The E3 ubiquitin ligase Triad3A negatively regulates the RIG-I/MAVS signaling pathway by targeting TRAF3 for degradation (PLoS pathogens, 2009)
- PMC5719392 — TRAF3 negatively regulates platelet activation and thrombosis (Scientific reports, 2017)
- PMC12358898 — Reduction of TRAF3 by heterozygosity or aging impacts B cell function (Proceedings of the National Academy of Sciences of the United States of America, 2025)