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- Table of Contents
THEM4 · Western blot design guide
THEM4 Western Blot Planning Guide
Plan a THEM4 Western blot around the catalog-observed 27.1 kDa band, image-backed A07814-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
THEM4 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 | 27.1 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Small intestine | |
| Negative control | Cerebral cortex |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 27.1 kDa |
| ⓘ | Localization | Cell membrane / Cell projection, ruffle membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated THEM4 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Pancreas |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A07814-1 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 THEM4 Western Blot Band Size?
Use the product-observed 27.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 27.1 kDa | Matches the authoritative product WB observation. |
| 27.1 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 THEM4 appearancePlan around 27.1 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 27.1 kDa; use this as the primary experimental expectation. |
| Calculated mass | 27.1 kDa from UniProt Q5T1C6; 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 A07814-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 27.1 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 THEM4 band
A07814-1 · THEM4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for THEM4 Western blot
🧪Use Small intestine as the first positive-control candidate and Cerebral cortex as the HPA Not detected negative candidate.
Positive control: Small intestine (High)
Negative control: Cerebral cortex (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for THEM4
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 |
|---|---|---|---|---|
| Small intestine | Reported tissue cells | High | Protein (HPA) | HPA → |
| Stomach | Reported tissue cells | High | Protein (HPA) | HPA → |
| Pancreas | Reported tissue cells | High | Protein (HPA) | HPA → |
| Placenta | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced THEM4 Western Blot Tips
Deeper troubleshooting and optimisation questions for THEM4, answered from its protein features.
Which band should guide the blot?
Use 27.1 kDa, the observation attached to the authoritative A07814-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 27.1 kDa expectation.
Which positive control should I start with?
Start with Small intestine, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Cerebral cortex 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 THEM4 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A07814-1 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
THEM4 Western Blot Reagents
Human-reactive THEM4 Western blot reagents with authoritative product imagery.
Real WB data
Anti-THEM4 Antibody Picoband®
Cat # A07814-1
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 Q5T1C6
- Human Protein Atlas — THEM4
- A07814-1 product record
- PMC8267908 — Exosomal miR-183-5p Shuttled by M2 Polarized Tumor-Associated Macrophage Promotes the Development of Colon Cancer via Targeting THEM4 Mediated PI3K/AKT and NF-κB Pathways (Frontiers in oncology, 2021)
- PMC12798084 — A mitochondria-related gene-based signature predicts pancreatic ductal adenocarcinoma clinical outcome and revealed CAMK2A/THEM4 regulates progression phenotypes and mitophagy in vivo and in vitro (Journal of translational medicine, 2025)