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- Table of Contents
CYP27B1 · Western blot design guide
CYP27B1 Western Blot Planning Guide
Plan a CYP27B1 Western blot around the catalog-observed 56.5 kDa band, image-backed A01370 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
CYP27B1 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 | 56.5 kDa observed | |
| Observed band | 56.5 kDa observed | |
| Gel | 12-15% | |
| Positive control | No defensible HPA tissue; use knockout/knockdown | |
| Negative control | Use knockout/knockdown or orthogonal negative |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 56.5 kDa |
| ⓘ | Localization | Mitochondrion membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated CYP27B1 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Known-positive human lysate |
| 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 | A01370 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 CYP27B1 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 CYP27B1 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 O15528; 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 A01370. |
| 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 CYP27B1 band
A01370 · CYP27B1Observed 56.5 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CYP27B1 Western blot
🧪No defensible HPA positive control was available; do not invent one.
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for CYP27B1
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 |
|---|
Undetected expression · recommended negative controls
The Human Protein Atlas exposes no structured IHC tissue profile for CYP27B1, so tissue-level positive/negative controls cannot be listed here. Use a documented CYP27B1-positive lysate (recombinant CYP27B1 or a known-expressing line) as the positive control and a CYP27B1 knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced CYP27B1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CYP27B1, answered from its protein features.
Which band should guide the blot?
Use 56.5 kDa, the observation attached to the authoritative A01370 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?
No defensible positive tissue was supplied; do not invent one.
Which negative control is defensible?
Use a target knockdown/knockout control when no HPA Not detected tissue is available.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for CYP27B1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01370 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
CYP27B1 Western Blot Reagents
Human/Mouse/Rat-reactive CYP27B1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-CYP27B1 Antibody Picoband®
Cat # A01370
Real WB data
Anti-CYP27B1 Antibody Picoband®
Cat # PB9548
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.