UCP2 · Western blot design guide

UCP2 Western Blot Planning Guide

Plan an UCP2 Western blot around the catalog-observed 33.2 kDa band, image-backed A02256-1 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 UCP2 (UCP2): expected band 33.2 kDa, antibody A02256-1, and guide-derived SDS-PAGE protocol steps
UCP2 Western blot protocol sheet — expected band 33.2 kDa, antibody A02256-1, controls and PMC citations. Open the full UCP2 WB guide →

UCP2 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 33.2 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Spleen
Negative control ⓘ Target knockdown/knockout
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 33.2 kDa
ⓘ Localization Mitochondrion inner membrane
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated UCP2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAppendix
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
PrimaryA02256-1 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 UCP2 Western Blot Band Size?

Use the product-observed 33.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
33.2 kDaMatches the authoritative product WB observation.
33.2 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected UCP2 appearancePlan around 33.2 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band33.2 kDa; use this as the primary experimental expectation.
Calculated mass33.2 kDa from UniProt P55851; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A02256-1.
Why is my band missing or off?
SituationLikely causeNext action
33.2 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 UCP2 Western blot

🧪Use Spleen as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Spleen (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for UCP2

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
Spleen Reported tissue cells High Protein (HPA) HPA →
Gallbladder Reported tissue cells High Protein (HPA) HPA →
Pancreas Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Section 3

Advanced UCP2 Western Blot Tips

Deeper troubleshooting and optimisation questions for UCP2, answered from its protein features.

Which band should guide the blot?
Use 33.2 kDa, the observation attached to the authoritative A02256-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 33.2 kDa expectation.
Which positive control should I start with?
Start with Spleen, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use a target knockdown/knockout control when no HPA Not detected tissue is available.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for UCP2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02256-1 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

UCP2 Western Blot Reagents

Human/Mouse/Rat-reactive UCP2 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for UCP2 using A02256-1; observed band 33.2 kDa
Anti-UCP2 Antibody Picoband®
Cat # A02256-1

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 P55851
  2. Human Protein Atlas — UCP2
  3. A02256-1 product record
  4. PMC7352594 — Brain Overexpression of Uncoupling Protein-2 (UCP2) Delays Renal Damage and Stroke Occurrence in Stroke-Prone Spontaneously Hypertensive Rats (International journal of molecular sciences, 2020)
  5. PMC13247729 — Pro- and Anti-Inflammatory Macrophages Adjust UCP2 Protein Levels Based on Their Intrinsic Metabolism and Available Metabolites (European journal of immunology, 2026)