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- Table of Contents
NFAT5 · Western blot design guide
NFAT5 Western Blot Planning Guide
Plan a NFAT5 Western blot around the catalog-observed 165.8 kDa band, image-backed A01815-3 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NFAT5 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 | 165.8 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Bronchus | |
| Negative control | Lymph node |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 165.8 kDa |
| ⓘ | Localization | Nucleus / Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NFAT5 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bronchus |
| Gel % | 8-10% |
| Load | 20-30 µg total protein per lane |
| Transfer | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A01815-3 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 NFAT5 Western Blot Band Size?
Use the product-observed 165.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 165.8 kDa | Matches the authoritative product WB observation. |
| 165.8 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 NFAT5 appearancePlan around 165.8 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 165.8 kDa; use this as the primary experimental expectation. |
| Calculated mass | 165.8 kDa from UniProt O94916; 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 A01815-3. |
| Situation | Likely cause | Next action |
|---|---|---|
| 165.8 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 NFAT5 band
A01815-3 · NFAT5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NFAT5 Western blot
🧪Use Bronchus as the first positive-control candidate and Lymph node as the HPA Not detected negative candidate.
Positive control: Bronchus (High)
Negative control: Lymph node (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for NFAT5
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 |
|---|---|---|---|---|
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Nasopharynx | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adipose tissue | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced NFAT5 Western Blot Tips
Deeper troubleshooting and optimisation questions for NFAT5, answered from its protein features.
Which band should guide the blot?
Use 165.8 kDa, the observation attached to the authoritative A01815-3 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 165.8 kDa expectation.
Which positive control should I start with?
Start with Bronchus, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Lymph node 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 NFAT5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01815-3 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
NFAT5 Western Blot Reagents
Human/Mouse/Rat-reactive NFAT5 Western blot reagents with authoritative product imagery.
Real WB data
Anti-NFAT5 Antibody Picoband®
Cat # A01815-3
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 O94916
- Human Protein Atlas — NFAT5
- A01815-3 product record
- PMC3388090 — NFAT5 is activated by hypoxia: role in ischemia and reperfusion in the rat kidney (PloS one, 2012)
- PMC13203630 — NFAT5: A Metabolic Time Capsule Encoding the History of Paternal Metabolic Oxidative Stress Within the Male Reproductive Tract (Antioxidants (Basel, Switzerland), 2026)
- PMC10602047 — Direct ionic stress sensing and mitigation by the transcription factor NFAT5 (bioRxiv : the preprint server for biology, 2023)