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- Table of Contents
ALKBH5 · Western blot design guide
ALKBH5 Western Blot Planning Guide
Plan an ALKBH5 Western blot around the catalog-observed 44.3 kDa band, image-backed A03360 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
ALKBH5 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 | 44.3 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Adipose tissue | |
| Negative control | Soft tissue |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 44.3 kDa |
| ⓘ | Localization | Nucleus speckle / Nucleus, nucleoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated ALKBH5 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adipose tissue |
| 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 | A03360 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 ALKBH5 Western Blot Band Size?
Use the product-observed 44.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 44.3 kDa | Matches the authoritative product WB observation. |
| 44.3 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 ALKBH5 appearancePlan around 44.3 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 44.3 kDa; use this as the primary experimental expectation. |
| Calculated mass | 44.3 kDa from UniProt Q6P6C2; 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 A03360. |
| Situation | Likely cause | Next action |
|---|---|---|
| 44.3 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 ALKBH5 band
A03360 · ALKBH5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ALKBH5 Western blot
🧪Use Adipose tissue as the first positive-control candidate and Soft tissue as the HPA Not detected negative candidate.
Positive control: Adipose tissue (High)
Negative control: Soft tissue (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for ALKBH5
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 → |
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Breast | 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 |
|---|---|---|---|---|
| Soft tissue | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
Section 3
Advanced ALKBH5 Western Blot Tips
Deeper troubleshooting and optimisation questions for ALKBH5, answered from its protein features.
Which band should guide the blot?
Use 44.3 kDa, the observation attached to the authoritative A03360 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 44.3 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 Soft tissue 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 ALKBH5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A03360 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
ALKBH5 Western Blot Reagents
Human-reactive ALKBH5 Western blot reagents with authoritative product imagery.
Real WB data
Anti-ALKBH5 Antibody Picoband®
Cat # A03360
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 Q6P6C2
- Human Protein Atlas — ALKBH5
- A03360 product record
- PMC7835694 — [ALKBH5 suppresses migration and invasion of human trophoblast cells by inhibiting epithelial-mesenchymal transition] (Nan fang yi ke da xue xue bao = Journal of Southern Medical University, 2020)
- PMC9814979 — Myocardial m6A regulators in postnatal development: effect of sex (Physiological research, 2022)
- PMC8755544 — RNA demethylase ALKBH5 promotes tumorigenesis in multiple myeloma via TRAF1-mediated activation of NF-κB and MAPK signaling pathways (Oncogene, 2022)