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- Table of Contents
SEMA3B · Western blot design guide
SEMA3B Western Blot Planning Guide
Plan a SEMA3B Western blot around the catalog-observed 83.1 kDa band, image-backed M06559 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SEMA3B Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | 83.1 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Adrenal gland | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 83.1 kDa |
| ⓘ | Localization | Secreted / Endoplasmic reticulum |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated SEMA3B Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| 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 | M06559 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 SEMA3B Western Blot Band Size?
Use the product-observed 83.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 83.1 kDa | Matches the authoritative product WB observation. |
| 83.1 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 SEMA3B appearancePlan around 83.1 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 83.1 kDa; use this as the primary experimental expectation. |
| Calculated mass | 83.1 kDa from UniProt Q13214; 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 M06559. |
| Situation | Likely cause | Next action |
|---|---|---|
| 83.1 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 SEMA3B band
M06559 · SEMA3B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SEMA3B Western blot
🧪Use Adrenal gland as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Adrenal gland (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for SEMA3B
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 → |
| Caudate | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced SEMA3B Western Blot Tips
Deeper troubleshooting and optimisation questions for SEMA3B, answered from its protein features.
Which band should guide the blot?
Use 83.1 kDa, the observation attached to the authoritative M06559 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 83.1 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 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 SEMA3B Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should M06559 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
SEMA3B Western Blot Reagents
Human/Mouse/Rat-reactive SEMA3B Western blot reagents with authoritative product imagery.
Real WB data
Anti-Semaphorin 3B/SEMA3B Antibody Picoband® (monoclonal, 9C4F7)
Cat # M06559
Real WB data
Anti-Semaphorin 3B/SEMA3B Antibody Picoband®
Cat # A06559-1
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 Q13214
- Human Protein Atlas — SEMA3B
- M06559 product record
- PMC2760388 — Semaphorin 3B inhibits the phosphatidylinositol 3-kinase/Akt pathway through neuropilin-1 in lung and breast cancer cells (Cancer research, 2008)
- PMC4511551 — Suppression of miR-221 inhibits glioma cells proliferation and invasion via targeting SEMA3B (Biological research, 2015)
