FHL2 · Western blot design guide

FHL2 Western Blot Planning Guide

Plan a FHL2 Western blot around the catalog-observed 32.2 kDa band, image-backed A02129-2 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 FHL2 (FHL2): expected band 32.2 kDa, antibody A02129-2, and guide-derived SDS-PAGE protocol steps
FHL2 Western blot protocol sheet — expected band 32.2 kDa, antibody A02129-2, controls and PMC citations. Open the full FHL2 WB guide →

FHL2 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 32.2 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Ovary
Negative control ⓘ Appendix
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 32.2 kDa
ⓘ Localization Cytoplasm / Nucleus
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated FHL2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateHeart muscle
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
PrimaryA02129-2 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 FHL2 Western Blot Band Size?

Use the product-observed 32.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
32.2 kDaMatches the authoritative product WB observation.
32.2 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected FHL2 appearancePlan around 32.2 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band32.2 kDa; use this as the primary experimental expectation.
Calculated mass32.2 kDa from UniProt Q14192; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A02129-2.
Why is my band missing or off?
SituationLikely causeNext action
32.2 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 FHL2 Western blot

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

HPA tissue expression evidence for FHL2

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
Ovary Reported tissue cells High Protein (HPA) HPA →
Kidney Reported tissue cells High Protein (HPA) HPA →
Heart muscle Reported tissue cells High Protein (HPA) HPA →
Cervix Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Bronchus Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced FHL2 Western Blot Tips

Deeper troubleshooting and optimisation questions for FHL2, answered from its protein features.

Which band should guide the blot?
Use 32.2 kDa, the observation attached to the authoritative A02129-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 32.2 kDa expectation.
Which positive control should I start with?
Start with Ovary, 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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for FHL2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02129-2 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

FHL2 Western Blot Reagents

Human/Mouse/Rat-reactive FHL2 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for FHL2 using A02129-2; observed band 32.2 kDa
Anti-FHL2 Antibody Picoband®
Cat # A02129-2

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 Q14192
  2. Human Protein Atlas — FHL2
  3. A02129-2 product record
  4. PMC12863675 — HCMV encoded UL84 hijacks FHL2 to suppress type I interferon production and enhance viral replication (PLoS pathogens, 2026)
  5. PMC8691187 — FHL2 interacts with EGFR to promote glioblastoma growth (Oncogene, 2018)
  6. PMC11460468 — FHL2 activates β-catenin/Wnt signaling by complexing with APC and TRIM63 in lung adenocarcinoma (Translational oncology, 2024)