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- Table of Contents
CXCR4 · Western blot design guide
CXCR4 Western Blot Planning Guide
Plan a CXCR4 Western blot around the catalog-observed 39.7 kDa band, image-backed A00031 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
CXCR4 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 | 39.7 kDa observed | |
| Observed band | 39.7 kDa observed | |
| Gel | 12-15% | |
| Positive control | No defensible HPA tissue; use knockout/knockdown | |
| Negative control | Use knockout/knockdown or orthogonal negative |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 39.7 kDa |
| ⓘ | Localization | Cell membrane / Cell junction |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated CXCR4 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Known-positive human lysate |
| 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 | A00031 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 CXCR4 Western Blot Band Size?
Use the product-observed 39.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 39.7 kDa | Matches the authoritative product WB observation. |
| 39.7 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 CXCR4 appearancePlan around 39.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 39.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 39.7 kDa from UniProt P61073; 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 A00031. |
| Situation | Likely cause | Next action |
|---|---|---|
| 39.7 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 CXCR4 band
A00031 · CXCR4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CXCR4 Western blot
🧪No defensible HPA positive control was available; do not invent one.
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for CXCR4
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 |
|---|
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced CXCR4 Western Blot Tips
Deeper troubleshooting and optimisation questions for CXCR4, answered from its protein features.
Which band should guide the blot?
Use 39.7 kDa, the observation attached to the authoritative A00031 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 39.7 kDa expectation.
Which positive control should I start with?
No defensible positive tissue was supplied; do not invent one.
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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for CXCR4 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00031 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
CXCR4 Western Blot Reagents
Human/Mouse/Rat-reactive CXCR4 Western blot reagents with authoritative product imagery.
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # A00031
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # A00031-2
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # A00031-3
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # A00031-4
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # PA1237
Real WB data
Anti-CXCR4 Antibody Picoband®
Cat # PA2081
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 P61073
- A00031 product record
- PMC4598861 — EZH2-mediated loss of miR-622 determines CXCR4 activation in hepatocellular carcinoma (Nature communications, 2015)
- PMC2645827 — Deubiquitination of CXCR4 by USP14 is critical for both CXCL12-induced CXCR4 degradation and chemotaxis but not ERK ativation (The Journal of biological chemistry, 2009)
- PMC3448054 — Inhibition of stromal CXCR4 impairs development of lung metastases (Cancer immunology, immunotherapy : CII, 2012)