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- Table of Contents
IDE · Western blot design guide
IDE Western Blot Planning Guide
Plan an IDE Western blot around the catalog-observed 118 kDa band, image-backed A01358-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
IDE 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 | 118 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Small intestine | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 118 kDa |
| ⓘ | Localization | Cytoplasm, cytosol / Cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Sample controls for IDE Western blot
🧪Use Small intestine as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.
Positive control: Small intestine (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 IDE
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart on each tissue page — 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 → |
| Heart muscle | Reported tissue cells | High | Protein (HPA) | HPA → |
| Skeletal muscle | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 1
What Is the Expected IDE Western Blot Band Size?
Use the product-observed 118 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 118 kDa | Matches the authoritative product WB observation. |
| 118 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 IDE appearancePlan around 118 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 118 kDa; use this as the primary experimental expectation. |
| Calculated mass | 118 kDa from UniProt P14735; 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 A01358-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 118 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 IDE band
A01358-2 · IDEObserved 118 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Section 2
Real Curated IDE Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Heart muscle |
| 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 | A01358-2 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 4
Advanced IDE Western Blot Tips
Deeper troubleshooting and optimisation questions for IDE, answered from its protein features.
Which band should guide the blot?
Use 118 kDa, the observation attached to the authoritative A01358-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 118 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 Bone marrow 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 IDE Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01358-2 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: PMC6675974, PMC4761760.
Boster reagents
IDE Western Blot Reagents
Human/Mouse/Rat-reactive IDE Western blot reagents with authoritative product imagery.
Real WB data
Anti-Insulin degrading enzyme/IDE Antibody Picoband®
Cat # A01358-2
Reactivity: Human, Mouse, Rat
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 P14735
- Human Protein Atlas — IDE
- A01358-2 product record
- PMC6675974 — Oxidative inactivation of amyloid beta-degrading proteases by cholesterol-enhanced mitochondrial stress.
- PMC4761760 — Genomic population structure of freshwater-resident and anadromous ide (Leuciscus idus) in north-western Europe.