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- Table of Contents
USP9X · Western blot design guide
USP9X Western Blot Planning Guide
Plan an USP9X Western blot around the catalog-observed 290.5 kDa band, image-backed A02594-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
USP9X 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 | 290.5 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 4-12% gradient | |
| Positive control | Colon | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 290.5 kDa |
| ⓘ | Localization | Cytoplasm, cytosol / Cell projection, growth cone |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated USP9X Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| Gel % | 4-12% gradient |
| Load | 20-30 µg total protein per lane |
| Transfer | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A02594-1 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 USP9X Western Blot Band Size?
Use the product-observed 290.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 290.5 kDa | Matches the authoritative product WB observation. |
| 290.5 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 USP9X appearancePlan around 290.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 290.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 290.5 kDa from UniProt Q93008; retain as context. |
| Gel selection | 4-12% gradient; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A02594-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 290.5 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 USP9X band
A02594-1 · USP9X
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for USP9X Western blot
🧪Use Colon as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Colon (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 USP9X
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 |
|---|---|---|---|---|
| Colon | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced USP9X Western Blot Tips
Deeper troubleshooting and optimisation questions for USP9X, answered from its protein features.
Which band should guide the blot?
Use 290.5 kDa, the observation attached to the authoritative A02594-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 290.5 kDa expectation.
Which positive control should I start with?
Start with Colon, 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 4-12% gradient consistently across the quick facts, protocol table, and poster.
What transfer method to use for USP9X Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02594-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC8191029, PMC5735260.
Boster reagents
USP9X Western Blot Reagents
Human/Mouse/Rat-reactive USP9X Western blot reagents with authoritative product imagery.
Real WB data
Anti-USP9X Antibody Picoband®
Cat # A02594-1
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 Q93008
- Human Protein Atlas — USP9X
- A02594-1 product record
- PMC8191029 — USP9X promotes apoptosis in cholangiocarcinoma by modulation expression of KIF1Bβ via deubiquitinating EGLN3.
- PMC5735260 — Usp9x Promotes Survival in Human Pancreatic Cancer and Its Inhibition Suppresses Pancreatic Ductal Adenocarcinoma In Vivo Tumor Growth.