KDM6B · Western blot design guide

KDM6B Western Blot Planning Guide

Plan a KDM6B Western blot around the catalog-observed 176.6 kDa band, image-backed A01309-1 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 KDM6B (KDM6B): expected band 176.6 kDa, antibody A01309-1, and guide-derived SDS-PAGE protocol steps
KDM6B Western blot protocol sheet — expected band 176.6 kDa, antibody A01309-1, controls and PMC citations. Open the full KDM6B WB guide →

KDM6B 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 176.6 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Cerebellum
Negative control ⓘ Soft tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 176.6 kDa
ⓘ Localization Nucleus
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse
Section 1

Real Curated KDM6B Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
Gel %8-10%
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA01309-1 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 2

What Is the Expected KDM6B Western Blot Band Size?

Use the product-observed 176.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
176.6 kDaMatches the authoritative product WB observation.
176.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected KDM6B appearancePlan around 176.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band176.6 kDa; use this as the primary experimental expectation.
Calculated mass176.6 kDa from UniProt O15054; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A01309-1.
Why is my band missing or off?
SituationLikely causeNext action
176.6 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 KDM6B Western blot

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

HPA tissue expression evidence for KDM6B

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
Cerebellum Reported tissue cells High Protein (HPA) HPA →
Colon Reported tissue cells High Protein (HPA) HPA →
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Pancreas Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced KDM6B Western Blot Tips

Deeper troubleshooting and optimisation questions for KDM6B, answered from its protein features.

Which band should guide the blot?
Use 176.6 kDa, the observation attached to the authoritative A01309-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 176.6 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Soft tissue 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 KDM6B Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01309-1 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

KDM6B Western Blot Reagents

Human/Mouse-reactive KDM6B Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for KDM6B using A01309-1; observed band 176.6 kDa
Anti-KDM6B/JMJD3 Picoband® Antibody
Cat # A01309-1

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

  1. UniProt O15054
  2. Human Protein Atlas — KDM6B
  3. A01309-1 product record
  4. PMC7863295 — Exosomal microRNA-22-3p alleviates cerebral ischemic injury by modulating KDM6B/BMP2/BMF axis (Stem cell research & therapy, 2021)
  5. PMC6328563 — Histone demethylase KDM6B has an anti-tumorigenic function in neuroblastoma by promoting differentiation (Oncogenesis, 2019)
  6. PMC5681448 — KDM6B modulates MAPK pathway mediating multiple myeloma cell growth and survival (Leukemia, 2017)