FLNA 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 280.7 kDa observed
Observed band 280.7 kDa observed
Gel 4-12% gradient
Positive control ⓘ Lung
Negative control ⓘ Bronchus
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass280.7 kDa
ⓘLocalizationCytoplasm, cell cortex / Cytoplasm, cytoskeleton
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Mouse / Rat
Section 1

Real Curated FLNA Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateBone marrow
Gel %4-12% gradient
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryM00502-2 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 FLNA Western Blot Band Size?

Use the product-observed 280.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
280.7 kDaMatches the authoritative product WB observation.
280.7 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected FLNA appearancePlan around 280.7 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band280.7 kDa; use this as the primary experimental expectation.
Calculated mass280.7 kDa from UniProt P21333; retain as context.
Gel selection4-12% gradient; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with M00502-2.
Why is my band missing or off?
SituationLikely causeNext action
280.7 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 FLNA Western blot

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

HPA tissue expression evidence for FLNA

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
Lung Reported tissue cells High Protein (HPA) HPA →
Ovary Reported tissue cells High Protein (HPA) HPA →
Kidney Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source
Bronchus Reported tissue cells Not detected Protein (HPA) HPA →
Caudate Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced FLNA Western Blot Tips

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

Which band should guide the blot?
Use 280.7 kDa, the observation attached to the authoritative M00502-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 280.7 kDa expectation.
Which positive control should I start with?
Start with Lung, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Bronchus as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 4-12% gradient consistently across the quick facts, protocol table, and poster.
What transfer method to use for FLNA Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should M00502-2 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

FLNA Western Blot Reagents

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

Real WB dataWestern blot validation image for FLNA using M00502-2; observed band 280.7 kDa
Anti-Filamin A/FLNA Antibody Picoband® (monoclonal, 3F8)
Cat # M00502-2

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.