PYROXD1 · Western blot design guide

PYROXD1 Western Blot Planning Guide

Plan a PYROXD1 Western blot around the catalog-observed 55.8 kDa band, image-backed A17214-3 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for PYROXD1 (PYROXD1): expected band 55.8 kDa, antibody A17214-3, and guide-derived SDS-PAGE protocol steps
PYROXD1 Western blot protocol sheet — expected band 55.8 kDa, antibody A17214-3, controls and PMC citations. Open the full PYROXD1 WB guide →

PYROXD1 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 55.8 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Adipose tissue
Negative control ⓘ Spleen
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 55.8 kDa
ⓘ Localization Nucleus / Cytoplasm, cytosol
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated PYROXD1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdipose tissue
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
PrimaryA17214-3 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 2

What Is the Expected PYROXD1 Western Blot Band Size?

Use the product-observed 55.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
55.8 kDaMatches the authoritative product WB observation.
55.8 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected PYROXD1 appearancePlan around 55.8 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band55.8 kDa; use this as the primary experimental expectation.
Calculated mass55.8 kDa from UniProt Q8WU10; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A17214-3.
Why is my band missing or off?
SituationLikely causeNext action
55.8 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.

Sample controls for PYROXD1 Western blot

🧪Use Adipose tissue as the first positive-control candidate and Spleen as the HPA Not detected negative candidate.
Positive control: Adipose tissue (Medium)
Negative control: Spleen (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for PYROXD1

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

TissueCell typeLevelEvidenceSource
Adipose tissue Reported tissue cells Medium Protein (HPA) HPA →
Breast Reported tissue cells Medium Protein (HPA) HPA →
Adrenal gland Reported tissue cells Medium Protein (HPA) HPA →
Appendix Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Spleen Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced PYROXD1 Western Blot Tips

Deeper troubleshooting and optimisation questions for PYROXD1, answered from its protein features.

Which band should guide the blot?
Use 55.8 kDa, the observation attached to the authoritative A17214-3 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 55.8 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Spleen 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 PYROXD1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A17214-3 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

PYROXD1 Western Blot Reagents

Human/Mouse/Rat-reactive PYROXD1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for PYROXD1 using A17214-3; observed band 55.8 kDa
Anti-PYROXD1 Antibody Picoband®
Cat # A17214-3
Real WB data Western blot validation image for PYROXD1 using A17214-4; observed band 55.8 kDa
Anti-PYROXD1 Antibody Picoband®
Cat # A17214-4

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

  1. UniProt Q8WU10
  2. Human Protein Atlas — PYROXD1
  3. A17214-3 product record
  4. PMC9637080 — Crosstalk between ROCK1 and PYROXD1 regulates CAFs activation and promotes laryngeal squamous cell carcinoma metastasis (Discover oncology, 2022)
  5. PMC13059403 — Pyroxd1 is essential for murine viability with the homozygous N155S recurrent variant linked to myopathy, muscle hypotrophy and osteopenia (Acta neuropathologica communications, 2026)
  6. PMC10244218 — Connective tissue presentation in two families expands the phenotypic spectrum of PYROXD1 disorders (Human molecular genetics, 2023)