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- Table of Contents
NEDD9 · Western blot design guide
NEDD9 Western Blot Planning Guide
Plan a NEDD9 Western blot around the catalog-observed 92.9 kDa band, image-backed PB9289 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NEDD9 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 | 92.9 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Adipose tissue | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 92.9 kDa |
| ⓘ | Localization | Cytoplasm, cell cortex / Nucleus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NEDD9 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adipose tissue |
| Gel % | 8-10% |
| 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 | PB9289 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 NEDD9 Western Blot Band Size?
Use the product-observed 92.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 92.9 kDa | Matches the authoritative product WB observation. |
| 92.9 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 NEDD9 appearancePlan around 92.9 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 92.9 kDa; use this as the primary experimental expectation. |
| Calculated mass | 92.9 kDa from UniProt Q14511; retain as context. |
| Gel selection | 8-10%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with PB9289. |
| Situation | Likely cause | Next action |
|---|---|---|
| 92.9 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 NEDD9 band
PB9289 · NEDD9
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NEDD9 Western blot
🧪Use Adipose tissue as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Adipose tissue (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for NEDD9
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 |
|---|---|---|---|---|
| Adipose tissue | Reported tissue cells | High | Protein (HPA) | HPA → |
| Gallbladder | Reported tissue cells | High | Protein (HPA) | HPA → |
| Caudate | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | High | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced NEDD9 Western Blot Tips
Deeper troubleshooting and optimisation questions for NEDD9, answered from its protein features.
Which band should guide the blot?
Use 92.9 kDa, the observation attached to the authoritative PB9289 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 92.9 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, 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 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for NEDD9 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9289 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
NEDD9 Western Blot Reagents
Human/Mouse/Rat-reactive NEDD9 Western blot reagents with authoritative product imagery.
Real WB data
Anti-HEF1/NEDD9 Antibody Picoband®
Cat # PB9289
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 Q14511
- Human Protein Atlas — NEDD9
- PB9289 product record
- PMC3145662 — NEDD9 is a positive regulator of epithelial-mesenchymal transition and promotes invasion in aggressive breast cancer (PloS one, 2011)
- PMC6119087 — NEDD9 promotes oncogenic signaling, a stem/mesenchymal gene signature, and aggressive ovarian cancer growth in mice (Oncogene, 2018)
- PMC6458266 — NEDD9 Facilitates Hypoxia-Induced Gastric Cancer Cell Migration via MICAL1 Related Rac1 Activation (Frontiers in pharmacology, 2019)