TLR3 · Western blot design guide

TLR3 Western Blot Planning Guide

Plan a TLR3 Western blot around the catalog-observed 103.8 kDa band, image-backed A00197 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 TLR3 (TLR3): expected band 103.8 kDa, antibody A00197, and guide-derived SDS-PAGE protocol steps
TLR3 Western blot protocol sheet — expected band 103.8 kDa, antibody A00197, controls and PMC citations. Open the full TLR3 WB guide →

TLR3 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 103.8 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Testis
Negative control ⓘ Appendix
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 103.8 kDa
ⓘ Localization Endoplasmic reticulum membrane / Endosome membrane
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated TLR3 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateTestis
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
PrimaryA00197 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 TLR3 Western Blot Band Size?

Use the product-observed 103.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
103.8 kDaMatches the authoritative product WB observation.
103.8 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected TLR3 appearancePlan around 103.8 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band103.8 kDa; use this as the primary experimental expectation.
Calculated mass103.8 kDa from UniProt O15455; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00197.
Why is my band missing or off?
SituationLikely causeNext action
103.8 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 TLR3 Western blot

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

HPA tissue expression evidence for TLR3

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
Testis Reported tissue cells High Protein (HPA) HPA →
Adrenal gland Reported tissue cells Medium Protein (HPA) HPA →
Colon Reported tissue cells Medium Protein (HPA) HPA →
Cerebellum Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced TLR3 Western Blot Tips

Deeper troubleshooting and optimisation questions for TLR3, answered from its protein features.

Which band should guide the blot?
Use 103.8 kDa, the observation attached to the authoritative A00197 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 103.8 kDa expectation.
Which positive control should I start with?
Start with Testis, 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 TLR3 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00197 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

TLR3 Western Blot Reagents

Human/Mouse/Rat-reactive TLR3 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for TLR3 using A00197; observed band 103.8 kDa
Anti-TLR3 Antibody Picoband®
Cat # A00197

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 O15455
  2. Human Protein Atlas — TLR3
  3. A00197 product record
  4. PMC3184062 — Differential toll-like receptor 3 (TLR3) expression and apoptotic response to TLR3 agonist in human neuroblastoma cells (Journal of biomedical science, 2011)
  5. PMC12804839 — Ubiquitination by HRD1 is essential for TLR3 trafficking and its innate immune signaling (Nature communications, 2025)
  6. PMC3978458 — MicroRNA-26a negatively regulates toll-like receptor 3 expression of rat macrophages and ameliorates pristane induced arthritis in rats (Arthritis research & therapy, 2014)