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- Table of Contents
NUMB · Western blot design guide
NUMB Western Blot Planning Guide
Plan a NUMB Western blot around the catalog-observed 70.8 kDa band, image-backed A01206-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NUMB 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 | 70.8 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Breast | |
| Negative control | Lymph node |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 70.8 kDa |
| ⓘ | Localization | Cell membrane / Endosome membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NUMB Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| 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 | A01206-2 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 NUMB Western Blot Band Size?
Use the product-observed 70.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 70.8 kDa | Matches the authoritative product WB observation. |
| 70.8 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 NUMB appearancePlan around 70.8 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 70.8 kDa; use this as the primary experimental expectation. |
| Calculated mass | 70.8 kDa from UniProt P49757; 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 A01206-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 70.8 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 NUMB band
A01206-2 · NUMB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NUMB Western blot
🧪Use Breast as the first positive-control candidate and Lymph node as the HPA Not detected negative candidate.
Positive control: Breast (High)
Negative control: Lymph node (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for NUMB
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 |
|---|---|---|---|---|
| Breast | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | 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 NUMB Western Blot Tips
Deeper troubleshooting and optimisation questions for NUMB, answered from its protein features.
Which band should guide the blot?
Use 70.8 kDa, the observation attached to the authoritative A01206-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 70.8 kDa expectation.
Which positive control should I start with?
Start with Breast, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Lymph node as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for NUMB Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01206-2 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
NUMB Western Blot Reagents
Human/Mouse/Rat-reactive NUMB Western blot reagents with authoritative product imagery.
Real WB data
Anti-NUMB Antibody Picoband®
Cat # A01206-2
Real WB data
Anti-NUMB Antibody Picoband®
Cat # PB9301
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 P49757
- Human Protein Atlas — NUMB
- A01206-2 product record
- PMC3514095 — RNAi-mediated knock-down of Dab and Numb attenuate Aβ levels via γ-secretase mediated APP processing (Translational neurodegeneration, 2012)
- PMC9637490 — [Numb activates the mTORC1 signaling pathway in proximal tubular epithelial cells by upregulating V1G1 expression] (Nan fang yi ke da xue xue bao = Journal of Southern Medical University, 2022)
- PMC4565853 — The Expression and Function of NUMB in Endometrial Cancer and the Interaction with HDM2 and P53 (Journal of Cancer, 2015)
