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- Table of Contents
NR4A1 · Western blot design guide
NR4A1 Western Blot Planning Guide
Plan a NR4A1 Western blot around the catalog-observed 64.5 kDa band, image-backed PA1676 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NR4A1 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 | Adrenal gland | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 64.5 kDa |
| ⓘ | Localization | Nucleus / Cytoplasm, cytosol |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NR4A1 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, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | PA1676 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 NR4A1 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 NR4A1 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 P22736; 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 PA1676. |
| 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 NR4A1 band
PA1676 · NR4A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NR4A1 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 NR4A1
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 → |
| Testis | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cervix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced NR4A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NR4A1, answered from its protein features.
Which band should guide the blot?
Use 64.5 kDa, the observation attached to the authoritative PA1676 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 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 NR4A1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PA1676 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
NR4A1 Western Blot Reagents
Human/Mouse/Rat-reactive NR4A1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-NUR77/NR4A1 Antibody Picoband®
Cat # PA1676
Real WB data
Anti-NUR77/NR4A1 Antibody Picoband®
Cat # PB9766
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 P22736
- Human Protein Atlas — NR4A1
- PA1676 product record
- PMC12936828 — Estrogen upregulates NR4A1 to counter TGF beta induced pulmonary fibrosis therapeutic insights for IPF (iScience, 2026)
- PMC12957799 — Age-dependent elevation of Nr4a1 attenuates PI3K/AKT/GSK3β pathway and mediates tau hyperphosphorylation and cognitive impairments (Journal of advanced research, 2026)
- PMC12678413 — Bruceine A protects nuclear receptor 4A1 from ubiquitin-degradation to alleviate mesangial proliferative glomerulonephritis (Signal transduction and targeted therapy, 2025)