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- Table of Contents
NPC2 · Western blot design guide
NPC2 Western Blot Planning Guide
Plan a NPC2 Western blot around the catalog-observed 16.6 kDa band, image-backed A01582-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NPC2 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 | 16.6 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 15% | |
| Positive control | Lung | |
| Negative control | Cerebral cortex |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 16.6 kDa |
| ⓘ | Localization | Secreted / Endoplasmic reticulum |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse |
Section 1
Real Curated NPC2 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Epididymis |
| Gel % | 15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.2 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A01582-2 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 NPC2 Western Blot Band Size?
Use the product-observed 16.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 16.6 kDa | Matches the authoritative product WB observation. |
| 16.6 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 NPC2 appearancePlan around 16.6 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 16.6 kDa; use this as the primary experimental expectation. |
| Calculated mass | 16.6 kDa from UniProt P61916; retain as context. |
| Gel selection | 15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A01582-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 16.6 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 NPC2 band
A01582-2 · NPC2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NPC2 Western blot
🧪Use Lung as the first positive-control candidate and Cerebral cortex as the HPA Not detected negative candidate.
Positive control: Lung (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 NPC2
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 |
|---|---|---|---|---|
| Lung | Reported tissue cells | High | Protein (HPA) | HPA → |
| Testis | Reported tissue cells | High | Protein (HPA) | HPA → |
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Epididymis | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced NPC2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NPC2, answered from its protein features.
Which band should guide the blot?
Use 16.6 kDa, the observation attached to the authoritative A01582-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 16.6 kDa expectation.
Which positive control should I start with?
Start with Lung, 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 15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for NPC2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01582-2 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents
NPC2 Western Blot Reagents
Human/Mouse-reactive NPC2 Western blot reagents with authoritative product imagery.
Real WB data
Anti-Niemann Pick C2/NPC2 Antibody Picoband®
Cat # A01582-2
Real WB data
Anti-Niemann Pick C2/NPC2 Picoband® Antibody
Cat # A01582-3
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 P61916
- Human Protein Atlas — NPC2
- A01582-2 product record
- PMC12274260 — ALKBH5-mediated NPC2 mRNA m(6)A demethylation promotes resistance to oxaliplatin in colorectal cancer (Functional & integrative genomics, 2025)
- PMC6032364 — Niemann-Pick Type C2 Protein Regulates Free Cholesterol Accumulation and Influences Hepatic Stellate Cell Proliferation and Mitochondrial Respiration Function (International journal of molecular sciences, 2018)
- PMC4964497 — Niemann-Pick Type C2 Protein Mediates Hepatic Stellate Cells Activation by Regulating Free Cholesterol Accumulation (International journal of molecular sciences, 2016)