UBA1 · Western blot design guide

UBA1 Western Blot Planning Guide

Plan an UBA1 Western blot around the catalog-observed 117.8 kDa band, image-backed PB9904 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 UBA1 (UBA1): expected band 117.8 kDa, antibody PB9904, and guide-derived SDS-PAGE protocol steps
UBA1 Western blot protocol sheet — expected band 117.8 kDa, antibody PB9904, controls and PMC citations. Open the full UBA1 WB guide →

UBA1 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 117.8 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Cervix
Negative control ⓘ Soft tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 117.8 kDa
ⓘ Localization Cytoplasm / Mitochondrion
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated UBA1 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
PrimaryPB9904 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 UBA1 Western Blot Band Size?

Use the product-observed 117.8 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
117.8 kDaMatches the authoritative product WB observation.
117.8 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected UBA1 appearancePlan around 117.8 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band117.8 kDa; use this as the primary experimental expectation.
Calculated mass117.8 kDa from UniProt P22314; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with PB9904.
Why is my band missing or off?
SituationLikely causeNext action
117.8 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 UBA1 Western blot

🧪Use Cervix as the first positive-control candidate and Soft tissue as the HPA Not detected negative candidate.
Positive control: Cervix (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 UBA1

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
Cervix Reported tissue cells High Protein (HPA) HPA →
Cerebellum Reported tissue cells High Protein (HPA) HPA →
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Bronchus Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced UBA1 Western Blot Tips

Deeper troubleshooting and optimisation questions for UBA1, answered from its protein features.

Which band should guide the blot?
Use 117.8 kDa, the observation attached to the authoritative PB9904 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 117.8 kDa expectation.
Which positive control should I start with?
Start with Cervix, 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 UBA1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9904 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

UBA1 Western Blot Reagents

Human/Mouse/Rat-reactive UBA1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for UBA1 using PB9904; observed band 117.8 kDa
Anti-E1 Ubiquitin Activating Enzyme/UBA1 Antibody Picoband®
Cat # PB9904

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 P22314
  2. Human Protein Atlas — UBA1
  3. PB9904 product record
  4. PMC12310546 — Characterization of E1 enzyme dependencies in mutant-UBA1 human cells reveals UBA6 as a novel therapeutic target in VEXAS syndrome (Leukemia, 2025)
  5. PMC10592724 — Shared and Distinct Mechanisms of UBA1 Inactivation Across Different Diseases (bioRxiv : the preprint server for biology, 2023)
  6. PMC11099125 — Shared and distinct mechanisms of UBA1 inactivation across different diseases (The EMBO journal, 2024)