This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
OTOF · Western blot design guide
OTOF Western Blot Planning Guide
Plan an OTOF Western blot around the catalog-observed 226.8 kDa band, image-backed PB9616 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
OTOF 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 | 226.8 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 4-12% gradient | |
| Positive control | Cerebellum | |
| Negative control | Appendix |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 226.8 kDa |
| ⓘ | Localization | Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane / Basolateral cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Mouse / Rat |
Section 1
Real Curated OTOF Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Cerebellum |
| 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 | PB9616 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 OTOF Western Blot Band Size?
Use the product-observed 226.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 226.8 kDa | Matches the authoritative product WB observation. |
| 226.8 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 OTOF appearancePlan around 226.8 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 226.8 kDa; use this as the primary experimental expectation. |
| Calculated mass | 226.8 kDa from UniProt Q9HC10; 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 PB9616. |
| Situation | Likely cause | Next action |
|---|---|---|
| 226.8 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 OTOF band
PB9616 · OTOF
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OTOF Western blot
🧪Use Cerebellum as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.
Positive control: Cerebellum (Medium)
Negative control: Appendix (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for OTOF
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 | Medium | Protein (HPA) | HPA → |
| Cerebral cortex | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Caudate | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | Low | Protein (HPA) | HPA → |
Section 3
Advanced OTOF Western Blot Tips
Deeper troubleshooting and optimisation questions for OTOF, answered from its protein features.
Which band should guide the blot?
Use 226.8 kDa, the observation attached to the authoritative PB9616 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 226.8 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Appendix 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 OTOF Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9616 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: PMC10776841, PMC9426595, PMC10797419.
Boster reagents
OTOF Western Blot Reagents
Mouse/Rat-reactive OTOF Western blot reagents with authoritative product imagery.
Real WB data
Anti-Otoferlin Antibody Picoband®
Cat # PB9616
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 Q9HC10
- Human Protein Atlas — OTOF
- PB9616 product record
- PMC10776841 — Slc20a1 and Slc20a2 regulate neuronal plasticity and cognition independently of their phosphate transport ability.
- PMC9426595 — Membrane Protein OTOF Is a Type I Interferon-Induced Entry Inhibitor of HIV-1 in Macrophages.
- PMC10797419 — Preclinical Efficacy And Safety Evaluation of AAV-OTOF in DFNB9 Mouse Model And Nonhuman Primate.