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- Table of Contents
SPAG5 · Western blot design guide
SPAG5 Western Blot Planning Guide
Plan a SPAG5 Western blot around the catalog-observed 134.4 kDa band, image-backed A07062-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SPAG5 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 | 134.4 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Testis | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 134.4 kDa |
| ⓘ | Localization | Cytoplasm / Cytoplasm, cytoskeleton |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated SPAG5 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bone marrow |
| Gel % | 8-10% |
| Load | 20-30 µg total protein per lane |
| Transfer | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A07062-2 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 SPAG5 Western Blot Band Size?
Use the product-observed 134.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 134.4 kDa | Matches the authoritative product WB observation. |
| 134.4 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 SPAG5 appearancePlan around 134.4 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 134.4 kDa; use this as the primary experimental expectation. |
| Calculated mass | 134.4 kDa from UniProt Q96R06; 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 A07062-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 134.4 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 SPAG5 band
A07062-2 · SPAG5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SPAG5 Western blot
🧪Use Testis as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Testis (High)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for SPAG5
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 |
|---|---|---|---|---|
| Testis | Reported tissue cells | High | Protein (HPA) | HPA → |
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
| Tonsil | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced SPAG5 Western Blot Tips
Deeper troubleshooting and optimisation questions for SPAG5, answered from its protein features.
Which band should guide the blot?
Use 134.4 kDa, the observation attached to the authoritative A07062-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 134.4 kDa expectation.
Which positive control should I start with?
Start with Testis, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Heart muscle as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for SPAG5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A07062-2 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC10433440, PMC6154423, PMC4669841.
Boster reagents
SPAG5 Western Blot Reagents
Human-reactive SPAG5 Western blot reagents with authoritative product imagery.
Real WB data
Anti-Astrin/Deepest/SPAG5 Antibody Picoband®
Cat # A07062-2
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 Q96R06
- Human Protein Atlas — SPAG5
- A07062-2 product record
- PMC10433440 — SPAG5, the upstream protein of Wnt and the target of curcumin, inhibits hepatocellular carcinoma.
- PMC6154423 — SPAG5 promotes hepatocellular carcinoma progression by downregulating SCARA5 through modifying β-catenin degradation.
- PMC4669841 — SPAG5 upregulation predicts poor prognosis in cervical cancer patients and alters sensitivity to taxol treatment via the mTOR signaling pathway.