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- Table of Contents
LAT · Western blot design guide
LAT Western Blot Planning Guide
Plan a LAT Western blot around the catalog-observed 27.9 kDa band, image-backed A01654-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
LAT 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 | 27.9 kDa observed | |
| Observed band | 27.9 kDa observed | |
| Gel | 12-15% | |
| Positive control | Spleen | |
| Negative control | Bronchus |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 27.9 kDa |
| ⓘ | Localization | Cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human |
Section 1
Real Curated LAT Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A01654-1 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 LAT Western Blot Band Size?
Use the product-observed 27.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 27.9 kDa | Matches the authoritative product WB observation. |
| 27.9 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 LAT appearancePlan around 27.9 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 27.9 kDa; use this as the primary experimental expectation. |
| Calculated mass | 27.9 kDa from UniProt O43561; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A01654-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 27.9 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 LAT band
A01654-1 · LAT
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LAT Western blot
🧪Use Spleen as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Spleen (High)
Negative control: Bronchus (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for LAT
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 |
|---|---|---|---|---|
| Spleen | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Lymph node | Reported tissue cells | High | Protein (HPA) | HPA → |
| Placenta | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced LAT Western Blot Tips
Deeper troubleshooting and optimisation questions for LAT, answered from its protein features.
Which band should guide the blot?
Use 27.9 kDa, the observation attached to the authoritative A01654-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 27.9 kDa expectation.
Which positive control should I start with?
Start with Spleen, 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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for LAT Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01654-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents
LAT Western Blot Reagents
Human-reactive LAT Western blot reagents with authoritative product imagery.
Real WB data
Anti-LAT Antibody Picoband®
Cat # A01654-1
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 O43561
- Human Protein Atlas — LAT
- A01654-1 product record
- PMC8094118 — PLCγ1 promotes phase separation of T cell signaling components (The Journal of cell biology, 2021)
- PMC7915312 — A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling (Cells, 2021)