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- Table of Contents
NAE1 · Western blot design guide
NAE1 Western Blot Planning Guide
Plan a NAE1 Western blot around the catalog-observed 60.2 kDa band, image-backed PB9339 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NAE1 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 | 60.2 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Cerebellum | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 60.2 kDa |
| ⓘ | Localization | Cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NAE1 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 | PB9339 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 NAE1 Western Blot Band Size?
Use the product-observed 60.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 60.2 kDa | Matches the authoritative product WB observation. |
| 60.2 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 NAE1 appearancePlan around 60.2 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 60.2 kDa; use this as the primary experimental expectation. |
| Calculated mass | 60.2 kDa from UniProt Q13564; 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 PB9339. |
| Situation | Likely cause | Next action |
|---|---|---|
| 60.2 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 NAE1 band
PB9339 · NAE1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NAE1 Western blot
🧪Use Cerebellum as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Cerebellum (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for NAE1
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 |
|---|---|---|---|---|
| Cerebellum | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced NAE1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NAE1, answered from its protein features.
Which band should guide the blot?
Use 60.2 kDa, the observation attached to the authoritative PB9339 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 60.2 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use a target knockdown/knockout control when no HPA Not detected tissue is available.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for NAE1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9339 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
NAE1 Western Blot Reagents
Human/Mouse/Rat-reactive NAE1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-APPBP1/NAE1 Antibody Picoband®
Cat # PB9339
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 Q13564
- Human Protein Atlas — NAE1
- PB9339 product record
- PMC9830515 — Neddylation of HER2 Inhibits its Protein Degradation and promotes Breast Cancer Progression (International journal of biological sciences, 2023)
- PMC11228393 — RBM15 Protects From Myocardial Infarction by Stabilizing NAE1 (JACC. Basic to translational science, 2024)
- PMC12098688 — Neddylation of RhoA impairs its protein degradation and promotes renal interstitial fibrosis progression in diabetic nephropathy (Acta pharmacologica Sinica, 2025)