IL27 · Western blot design guide

IL27 Western Blot Planning Guide

Plan a IL27 Western blot around the catalog-observed 27.5 kDa band, image-backed A00857-1 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for IL27 (IL27): expected band 27.5 kDa observed, antibody A00857-1, and guide-derived SDS-PAGE protocol steps
IL27 Western blot protocol sheet — expected band 27.5 kDa observed, antibody A00857-1, controls and PMC citations. Open the full IL27 WB guide →

IL27 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 27.5 kDa observed
Observed band 27.5 kDa observed
Gel 12-15%
Positive control ⓘ Adipose tissue
Negative control ⓘ Adipose tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 27.5 kDa
ⓘ Localization Secreted
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human
Section 1

Real Curated IL27 Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysateAdipose tissue
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, short transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA00857-1 at datasheet starting dilution
Primary incubationOvernight at 4 °C with gentle agitation
SecondarySpecies-matched HRP conjugate at validated dilution
Wash3 × 5 min in TBST
DetectionChemiluminescent substrate
ExposureBracket exposures to avoid saturation
Section 2

What Is the Expected IL27 Western Blot Band Size?

Use the product-observed 27.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
27.5 kDaMatches the authoritative product WB observation.
27.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected IL27 appearancePlan around 27.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band27.5 kDa; use this as the primary experimental expectation.
Calculated mass27.5 kDa from UniProt Q8NEV9; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00857-1.
Why is my band missing or off?
SituationLikely causeNext action
27.5 kDaMatches the authoritative product WB observation.Confirm with orthogonal controls and the linked product record.
Additional bandMay reflect processing, modification, or non-specific signal.Run a dilution series and compare positive/negative controls.
Weak signalTarget abundance or transfer may be limiting.Verify transfer, increase positive-control abundance, and bracket exposure.

Sample controls for IL27 Western blot

🧪Use Adipose tissue as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Adipose tissue (Not detected)
Negative control: Adipose tissue (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for IL27

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

TissueCell typeLevelEvidenceSource

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Reported tissue cells Not detected Protein (HPA) HPA →
Adrenal gland Reported tissue cells Not detected Protein (HPA) HPA →
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced IL27 Western Blot Tips

Deeper troubleshooting and optimisation questions for IL27, answered from its protein features.

Which band should guide the blot?
Use 27.5 kDa, the observation attached to the authoritative A00857-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 27.5 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Adipose tissue 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 IL27 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00857-1 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

IL27 Western Blot Reagents

Human-reactive IL27 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for IL27 using A00857-1; observed band 27.5 kDa
Anti-IL27 Antibody Picoband®
Cat # A00857-1

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

  1. UniProt Q8NEV9
  2. Human Protein Atlas — IL27
  3. A00857-1 product record
  4. PMC9596272 — Interferon-γ Stimulates Interleukin-27 Derived from Dendritic Cells to Regulate Th9 Differentiation through STAT1/3 Pathway (Disease markers, 2022)