ANLN · Western blot design guide

ANLN Western Blot Planning Guide

Plan an ANLN Western blot around the catalog-observed 124.2 kDa band, image-backed A03997-1 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 ANLN (ANLN): expected band 124.2 kDa, antibody A03997-1, and guide-derived SDS-PAGE protocol steps
ANLN Western blot protocol sheet — expected band 124.2 kDa, antibody A03997-1, controls and PMC citations. Open the full ANLN WB guide →

ANLN 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 124.2 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Colon
Negative control ⓘ Epididymis
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 124.2 kDa
ⓘ Localization Nucleus / Cytoplasm, cytoskeleton
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated ANLN Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAppendix
Gel %8-10%
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA03997-1 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 ANLN Western Blot Band Size?

Use the product-observed 124.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
124.2 kDaMatches the authoritative product WB observation.
124.2 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ANLN appearancePlan around 124.2 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band124.2 kDa; use this as the primary experimental expectation.
Calculated mass124.2 kDa from UniProt Q9NQW6; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A03997-1.
Why is my band missing or off?
SituationLikely causeNext action
124.2 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 ANLN Western blot

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

HPA tissue expression evidence for ANLN

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
Colon Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →
Duodenum Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Epididymis Reported tissue cells Not detected Protein (HPA) HPA →
Heart muscle Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced ANLN Western Blot Tips

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

Which band should guide the blot?
Use 124.2 kDa, the observation attached to the authoritative A03997-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 124.2 kDa expectation.
Which positive control should I start with?
Start with Colon, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Epididymis 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 ANLN Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A03997-1 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

ANLN Western Blot Reagents

Human/Mouse/Rat-reactive ANLN Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for ANLN using A03997-1; observed band 124.2 kDa
Anti-Anillin/ANLN Antibody Picoband®
Cat # A03997-1

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 Q9NQW6
  2. Human Protein Atlas — ANLN
  3. A03997-1 product record
  4. PMC7891526 — Actin-binding protein Anillin promotes the progression of gastric cancer in vitro and in mice (Journal of clinical laboratory analysis, 2021)
  5. PMC9950447 — Targeting USP10 induces degradation of oncogenic ANLN in esophageal squamous cell carcinoma (Cell death and differentiation, 2023)
  6. PMC11834070 — ANLN promotes head and neck squamous cell carcinoma progression by upregulating PD-L1 via the ERK-MAPK pathway (iScience, 2025)