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- Table of Contents
IRF9 · Western blot design guide
IRF9 Western Blot Planning Guide
Plan an IRF9 Western blot around the catalog-observed 43.7 kDa band, image-backed A04485 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
IRF9 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 | 43.7 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Fallopian tube | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 43.7 kDa |
| ⓘ | Localization | Cytoplasm / Nucleus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated IRF9 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bronchus |
| 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 | A04485 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 IRF9 Western Blot Band Size?
Use the product-observed 43.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 43.7 kDa | Matches the authoritative product WB observation. |
| 43.7 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 IRF9 appearancePlan around 43.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 43.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 43.7 kDa from UniProt Q00978; 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 A04485. |
| Situation | Likely cause | Next action |
|---|---|---|
| 43.7 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 IRF9 band
A04485 · IRF9
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IRF9 Western blot
🧪Use Fallopian tube as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Fallopian tube (High)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for IRF9
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 |
|---|---|---|---|---|
| Fallopian tube | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Endometrium | Reported tissue cells | High | Protein (HPA) | HPA → |
| Epididymis | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced IRF9 Western Blot Tips
Deeper troubleshooting and optimisation questions for IRF9, answered from its protein features.
Which band should guide the blot?
Use 43.7 kDa, the observation attached to the authoritative A04485 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 43.7 kDa expectation.
Which positive control should I start with?
Start with Fallopian tube, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Heart muscle 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 IRF9 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04485 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
IRF9 Western Blot Reagents
Human-reactive IRF9 Western blot reagents with authoritative product imagery.
Real WB data
Anti-Interferon regulatory factor 9/IRF9 Antibody Picoband®
Cat # A04485
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 Q00978
- Human Protein Atlas — IRF9
- A04485 product record
- PMC8079643 — SARS-CoV-2 Spike Targets USP33-IRF9 Axis via Exosomal miR-148a to Activate Human Microglia (Frontiers in immunology, 2021)
- PMC6345781 — STAT3 is activated in multicellular spheroids of colon carcinoma cells and mediates expression of IRF9 and interferon stimulated genes (Scientific reports, 2019)
- PMC9250956 — [Effects of interferon regulatory factor 9 on the biological phenotypes in PML-RARα-induced promyelocytic leukemia] (Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi, 2022)