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 mass79.7 kDa
LocalizationPostsynaptic cell membrane / Cell membrane
Processing / PTMRecord-dependent
ReactivityHuman / Mouse / Rat

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 on each tissue page — 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 →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source
Heart muscle Reported tissue cells Not detected Protein (HPA) HPA →
Skeletal muscle Reported tissue cells Not detected Protein (HPA) HPA →
Section 1

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.

What am I looking at on my blot?
79.7 kDaMatches the authoritative product WB observation.
79.7 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected GPHN appearancePlan around 79.7 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band79.7 kDa; use this as the primary experimental expectation.
Calculated mass79.7 kDa from UniProt Q9NQX3; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A04560-1.
Why is my band missing or off?
SituationLikely causeNext action
79.7 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.
Section 2

Real Curated GPHN Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateCaudate
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA04560-1 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 4

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 dataWestern blot validation image for GPHN using A04560-1; observed band 79.7 kDa
Anti-Gephyrin/GPHN Antibody Picoband®
Cat # A04560-1
Reactivity: Human, Mouse, Rat, Monkey

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.