ACTR3 · Western blot design guide

ACTR3 Western Blot Planning Guide

Plan an ACTR3 Western blot around the catalog-observed 47.4 kDa band, image-backed A03423-2 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 ACTR3 (ACTR3): expected band 47.4 kDa, antibody A03423-2, and guide-derived SDS-PAGE protocol steps
ACTR3 Western blot protocol sheet — expected band 47.4 kDa, antibody A03423-2, controls and PMC citations. Open the full ACTR3 WB guide →

ACTR3 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 47.4 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Breast
Negative control ⓘ Soft tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 47.4 kDa
ⓘ Localization Cytoplasm, cytoskeleton / Cell projection
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated ACTR3 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateBone marrow
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA03423-2 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 ACTR3 Western Blot Band Size?

Use the product-observed 47.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
47.4 kDaMatches the authoritative product WB observation.
47.4 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ACTR3 appearancePlan around 47.4 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band47.4 kDa; use this as the primary experimental expectation.
Calculated mass47.4 kDa from UniProt P61158; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A03423-2.
Why is my band missing or off?
SituationLikely causeNext action
47.4 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 ACTR3 Western blot

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

HPA tissue expression evidence for ACTR3

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
Breast Reported tissue cells High Protein (HPA) HPA →
Kidney Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →
Lymph node Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Soft tissue Reported tissue cells Not detected Protein (HPA) HPA →
Parathyroid gland Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced ACTR3 Western Blot Tips

Deeper troubleshooting and optimisation questions for ACTR3, answered from its protein features.

Which band should guide the blot?
Use 47.4 kDa, the observation attached to the authoritative A03423-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 47.4 kDa expectation.
Which positive control should I start with?
Start with Breast, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Soft 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 ACTR3 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A03423-2 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

ACTR3 Western Blot Reagents

Human/Mouse/Rat-reactive ACTR3 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for ACTR3 using A03423-2; observed band 47.4 kDa
Anti-Arp3/ACTR3 Antibody Picoband®
Cat # A03423-2

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 P61158
  2. Human Protein Atlas — ACTR3
  3. A03423-2 product record
  4. PMC8539850 — Decreased exosome-delivered miR-486-5p is responsible for the peritoneal metastasis of gastric cancer cells by promoting EMT progress (World journal of surgical oncology, 2021)
  5. PMC7882613 — Induced Arp2/3 Complex Depletion Increases FMNL2/3 Formin Expression and Filopodia Formation (Frontiers in cell and developmental biology, 2021)