RNF2 · Western blot design guide

RNF2 Western Blot Planning Guide

Plan a RNF2 Western blot around the catalog-observed 37.7 kDa band, image-backed A01209-3 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 RNF2 (RNF2): expected band 37.7 kDa, antibody A01209-3, and guide-derived SDS-PAGE protocol steps
RNF2 Western blot protocol sheet — expected band 37.7 kDa, antibody A01209-3, controls and PMC citations. Open the full RNF2 WB guide →

RNF2 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 37.7 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Caudate
Negative control ⓘ Appendix
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 37.7 kDa
ⓘ Localization Nucleus / Cytoplasm
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human
Section 1

Real Curated RNF2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateBronchus
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
PrimaryA01209-3 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 RNF2 Western Blot Band Size?

Use the product-observed 37.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
37.7 kDaMatches the authoritative product WB observation.
37.7 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected RNF2 appearancePlan around 37.7 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band37.7 kDa; use this as the primary experimental expectation.
Calculated mass37.7 kDa from UniProt Q99496; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A01209-3.
Why is my band missing or off?
SituationLikely causeNext action
37.7 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.

Sample controls for RNF2 Western blot

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

HPA tissue expression evidence for RNF2

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
Caudate Reported tissue cells Medium Protein (HPA) HPA →
Liver Reported tissue cells Medium Protein (HPA) HPA →
Bronchus Reported tissue cells Medium Protein (HPA) HPA →
Duodenum Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Cerebral cortex Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced RNF2 Western Blot Tips

Deeper troubleshooting and optimisation questions for RNF2, answered from its protein features.

Which band should guide the blot?
Use 37.7 kDa, the observation attached to the authoritative A01209-3 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 37.7 kDa expectation.
Which positive control should I start with?
Start with Caudate, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Appendix 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 RNF2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01209-3 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

RNF2 Western Blot Reagents

Human-reactive RNF2 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for RNF2 using A01209-3; observed band 37.7 kDa
Anti-RING2/RING1B/RNF2 Antibody Picoband®
Cat # A01209-3

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 Q99496
  2. Human Protein Atlas — RNF2
  3. A01209-3 product record
  4. PMC9813066 — The ubiquitin ligase RNF2 stabilizes ERα and modulates breast cancer progression (Human cell, 2023)
  5. PMC3920452 — Knockdown of RNF2 induces apoptosis by regulating MDM2 and p53 stability (Oncogene, 2014)