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- Table of Contents
GPHN · Western blot design guide
GPHN Western Blot Planning Guide
Plan a GPHN Western blot around the catalog-observed 79.7 kDa band, image-backed A04560-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
GPHN 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 | 79.7 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Caudate | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 79.7 kDa |
| ⓘ | Localization | Postsynaptic cell membrane / Cell membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated GPHN Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Caudate |
| Gel % | 12-15% |
| 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 | A04560-1 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 GPHN Western Blot Band Size?
Use the product-observed 79.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 79.7 kDa | Matches the authoritative product WB observation. |
| 79.7 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 GPHN appearancePlan around 79.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 79.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 79.7 kDa from UniProt Q9NQX3; 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 A04560-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 79.7 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 GPHN band
A04560-1 · GPHN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GPHN Western blot
🧪Use Caudate as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Caudate (High)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for GPHN
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 |
|---|---|---|---|---|
| Caudate | Reported tissue cells | High | Protein (HPA) | HPA → |
| Hippocampus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebral cortex | Reported tissue cells | High | Protein (HPA) | HPA → |
| Endometrium | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced GPHN Western Blot Tips
Deeper troubleshooting and optimisation questions for GPHN, answered from its protein features.
Which band should guide the blot?
Use 79.7 kDa, the observation attached to the authoritative A04560-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 79.7 kDa expectation.
Which positive control should I start with?
Start with Caudate, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Heart muscle 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 GPHN Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04560-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC11549329, PMC10760056, PMC6599520.
Boster reagents
GPHN Western Blot Reagents
Human/Mouse/Rat-reactive GPHN Western blot reagents with authoritative product imagery.
Real WB data
Anti-Gephyrin/GPHN Antibody Picoband®
Cat # A04560-1
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 Q9NQX3
- Human Protein Atlas — GPHN
- A04560-1 product record
- PMC11549329 — miRNA-mediated control of gephyrin synthesis drives sustained inhibitory synaptic plasticity.
- PMC10760056 — miRNA-mediated control of gephyrin synthesis drives sustained inhibitory synaptic plasticity.
- PMC6599520 — Spatial patterning of excitatory and inhibitory neuropil territories during spinal circuit development.