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- Table of Contents
ACKR2 · Western blot design guide
ACKR2 Western Blot Planning Guide
Plan an ACKR2 Western blot around the catalog-observed 43.4 kDa band, image-backed A05491-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
ACKR2 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 | 43.4 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Fallopian tube | |
| Negative control | Appendix |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 43.4 kDa |
| ⓘ | Localization | Early endosome / Recycling endosome |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated ACKR2 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bronchus |
| 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 | A05491-2 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 ACKR2 Western Blot Band Size?
Use the product-observed 43.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 43.4 kDa | Matches the authoritative product WB observation. |
| 43.4 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 ACKR2 appearancePlan around 43.4 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 43.4 kDa; use this as the primary experimental expectation. |
| Calculated mass | 43.4 kDa from UniProt O00590; 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 A05491-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 43.4 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 ACKR2 band
A05491-2 · ACKR2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ACKR2 Western blot
🧪Use Fallopian tube as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.
Positive control: Fallopian tube (High)
Negative control: Appendix (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for ACKR2
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 |
|---|---|---|---|---|
| Fallopian tube | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Placenta | Reported tissue cells | High | Protein (HPA) | HPA → |
| Duodenum | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced ACKR2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ACKR2, answered from its protein features.
Which band should guide the blot?
Use 43.4 kDa, the observation attached to the authoritative A05491-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 43.4 kDa expectation.
Which positive control should I start with?
Start with Fallopian tube, 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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for ACKR2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A05491-2 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
ACKR2 Western Blot Reagents
Human-reactive ACKR2 Western blot reagents with authoritative product imagery.
Real WB data
Anti-D6/ACKR2 Antibody Picoband®
Cat # A05491-2
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 O00590
- Human Protein Atlas — ACKR2
- A05491-2 product record
- PMC4007430 — The N-terminal region of the atypical chemokine receptor ACKR2 is a key determinant of ligand binding (The Journal of biological chemistry, 2014)
- PMC5511891 — The chemokine scavenging receptor D6/ACKR2 is a target of miR-146a in thyroid cancer (Genes & cancer, 2017)
- PMC10000713 — Decoy Receptors Regulation by Resveratrol in Lipopolysaccharide-Activated Microglia (Cells, 2023)