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- Table of Contents
TLR4 · Western blot design guide
TLR4 Western Blot Planning Guide
Plan a TLR4 Western blot around the catalog-observed 95.7 kDa band, image-backed A00017-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
TLR4 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 | 95.7 kDa observed | |
| Observed band | 95.7 kDa observed | |
| Gel | 8-10% | |
| Positive control | Bone marrow | |
| Negative control | Appendix |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 95.7 kDa |
| ⓘ | Localization | Cell membrane / Early endosome |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated TLR4 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bone marrow |
| Gel % | 8-10% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A00017-2 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected TLR4 Western Blot Band Size?
Use the product-observed 95.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 95.7 kDa | Matches the authoritative product WB observation. |
| 95.7 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected TLR4 appearancePlan around 95.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 95.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 95.7 kDa from UniProt O00206; retain as context. |
| Gel selection | 8-10%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A00017-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 95.7 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing TLR4 band
A00017-2 · TLR4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TLR4 Western blot
🧪Use Bone marrow as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Bone marrow (High)
Negative control: Appendix (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for TLR4
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Reported tissue cells | High | Protein (HPA) | HPA → |
| Spleen | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Lymph node | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Low | Protein (HPA) | HPA → |
Section 3
Advanced TLR4 Western Blot Tips
Deeper troubleshooting and optimisation questions for TLR4, answered from its protein features.
Which band should guide the blot?
Use 95.7 kDa, the observation attached to the authoritative A00017-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 95.7 kDa expectation.
Which positive control should I start with?
Start with Bone marrow, 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 TLR4 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00017-2 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
TLR4 Western Blot Reagents
Human-reactive TLR4 Western blot reagents with authoritative product imagery.
Real WB data
Anti-TLR4 Antibody Picoband®
Cat # A00017-2
Real WB data
Anti-Toll-like receptor 4 TLR4 Antibody Picoband®
Cat # PA1484
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
- UniProt O00206
- Human Protein Atlas — TLR4
- A00017-2 product record
- PMC2675888 — Role of TLR4 tyrosine phosphorylation in signal transduction and endotoxin tolerance (The Journal of biological chemistry, 2007)