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- Table of Contents
SI · Western blot design guide
SI Western Blot Planning Guide
Plan a SI Western blot around the catalog-observed 209.5 kDa band, image-backed A04542-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SI 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 | 209.5 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 4-12% gradient | |
| Positive control | Small intestine | |
| Negative control | Bone marrow |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 209.5 kDa |
| ⓘ | Localization | Apical cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated SI Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Duodenum |
| Gel % | 4-12% gradient |
| 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 | A04542-1 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 SI Western Blot Band Size?
Use the product-observed 209.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 209.5 kDa | Matches the authoritative product WB observation. |
| 209.5 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 SI appearancePlan around 209.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 209.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 209.5 kDa from UniProt P14410; retain as context. |
| Gel selection | 4-12% gradient; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A04542-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 209.5 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 SI band
A04542-1 · SI
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SI 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 SI
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.
Section 3
Advanced SI Western Blot Tips
Deeper troubleshooting and optimisation questions for SI, answered from its protein features.
Which band should guide the blot?
Use 209.5 kDa, the observation attached to the authoritative A04542-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 209.5 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 4-12% gradient consistently across the quick facts, protocol table, and poster.
What transfer method to use for SI Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04542-1 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
SI Western Blot Reagents
Human/Mouse/Rat-reactive SI Western blot reagents with authoritative product imagery.
Real WB data
Anti-SI Antibody Picoband®
Cat # A04542-1
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 P14410
- Human Protein Atlas — SI
- A04542-1 product record
- PMC6123821 — Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation (Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 2018)