ACTN2 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 103.9 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control Heart muscle
Negative control Appendix
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass103.9 kDa
LocalizationCytoplasm, myofibril, sarcomere, Z line
Processing / PTMRecord-dependent
ReactivityHuman / Mouse / Rat

Sample controls for ACTN2 Western blot

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

HPA tissue expression evidence for ACTN2

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
Heart muscle Reported tissue cells High Protein (HPA) HPA →
Skeletal muscle Reported tissue cells High Protein (HPA) HPA →
Caudate Reported tissue cells Medium Protein (HPA) HPA →
Testis Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Section 1

What Is the Expected ACTN2 Western Blot Band Size?

Use the product-observed 103.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
103.9 kDaMatches the authoritative product WB observation.
103.9 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ACTN2 appearancePlan around 103.9 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band103.9 kDa; use this as the primary experimental expectation.
Calculated mass103.9 kDa from UniProt P35609; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A03673-1.
Why is my band missing or off?
SituationLikely causeNext action
103.9 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.
Section 2

Real Curated ACTN2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateHeart muscle
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
PrimaryA03673-1 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 4

Advanced ACTN2 Western Blot Tips

Deeper troubleshooting and optimisation questions for ACTN2, answered from its protein features.

Which band should guide the blot?
Use 103.9 kDa, the observation attached to the authoritative A03673-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 103.9 kDa expectation.
Which positive control should I start with?
Start with Heart muscle, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Appendix 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 ACTN2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A03673-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC9454684, PMC8692448.
Boster reagents

ACTN2 Western Blot Reagents

Human/Mouse/Rat-reactive ACTN2 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for ACTN2 using A03673-1; observed band 103.9 kDa
Anti-ACTN2 Antibody Picoband®
Cat # A03673-1
Reactivity: Human, Mouse, Rat

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.