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- Table of Contents
PIAS4 · Western blot design guide
PIAS4 Western Blot Planning Guide
Plan a PIAS4 Western blot around the catalog-observed 56.5 kDa band, image-backed PA2215 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
PIAS4 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 | 56.5 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Testis | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 56.5 kDa |
| ⓘ | Localization | Nucleus, PML body |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated PIAS4 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Testis |
| 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 | PA2215 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 PIAS4 Western Blot Band Size?
Use the product-observed 56.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 56.5 kDa | Matches the authoritative product WB observation. |
| 56.5 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 PIAS4 appearancePlan around 56.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 56.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 56.5 kDa from UniProt Q8N2W9; 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 PA2215. |
| Situation | Likely cause | Next action |
|---|---|---|
| 56.5 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 PIAS4 band
PA2215 · PIAS4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PIAS4 Western blot
🧪Use Testis as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.
Positive control: Testis (High)
Negative control: Bone marrow (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for PIAS4
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 → |
| Breast | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Low | Protein (HPA) | HPA → |
Section 3
Advanced PIAS4 Western Blot Tips
Deeper troubleshooting and optimisation questions for PIAS4, answered from its protein features.
Which band should guide the blot?
Use 56.5 kDa, the observation attached to the authoritative PA2215 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 56.5 kDa expectation.
Which positive control should I start with?
Start with Testis, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Bone marrow 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 PIAS4 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PA2215 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
PIAS4 Western Blot Reagents
Human/Mouse/Rat-reactive PIAS4 Western blot reagents with authoritative product imagery.
Real WB data
Anti-E3 SUMO-protein ligase PIAS4 Antibody Picoband®
Cat # PA2215
Real WB data
Anti-E3 SUMO-protein ligase PIAS4 Antibody Picoband®
Cat # PB10082
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 Q8N2W9
- Human Protein Atlas — PIAS4
- PA2215 product record
- PMC13139036 — PIAS4 inhibition induces cell cycle arrest and exhibits a synergistic effect in combination with CDK4/6 inhibitor in breast cancer treatment (Oncogene, 2026)
- PMC11954794 — Tanshinone IIA destabilizes SLC7A11 by regulating PIAS4-mediated SUMOylation of SLC7A11 through KDM1A, and promotes ferroptosis in breast cancer (Journal of advanced research, 2025)
- PMC3790182 — PIAS4 is an activator of hypoxia signalling via VHL suppression during growth of pancreatic cancer cells (British journal of cancer, 2013)