RGN · Western blot design guide

RGN Western Blot Planning Guide

Plan a RGN Western blot around the catalog-observed 33.3 kDa band, image-backed A05900-1 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 RGN (RGN): expected band 33.3 kDa, antibody A05900-1, and PMC-cited SDS-PAGE protocol steps
RGN Western blot protocol sheet — expected band 33.3 kDa, antibody A05900-1, controls and PMC citations. Open the full RGN WB guide →

RGN 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 33.3 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Adrenal gland
Negative control ⓘ Appendix
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 33.3 kDa
ⓘ Localization Cytoplasm
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated RGN Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
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
PrimaryA05900-1 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 RGN Western Blot Band Size?

Use the product-observed 33.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
33.3 kDaMatches the authoritative product WB observation.
33.3 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected RGN appearancePlan around 33.3 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band33.3 kDa; use this as the primary experimental expectation.
Calculated mass33.3 kDa from UniProt Q15493; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A05900-1.
Why is my band missing or off?
SituationLikely causeNext action
33.3 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 RGN Western blot

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

HPA tissue expression evidence for RGN

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
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Liver Reported tissue cells High Protein (HPA) HPA →
Pancreas 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 →
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced RGN Western Blot Tips

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

Which band should guide the blot?
Use 33.3 kDa, the observation attached to the authoritative A05900-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 33.3 kDa expectation.
Which positive control should I start with?
Start with Adrenal gland, 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 RGN Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A05900-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC12446341, PMC11031438.
Boster reagents

RGN Western Blot Reagents

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

Real WB data Western blot validation image for RGN using A05900-1; observed band 33.3 kDa
Anti-Regucalcin/RGN Antibody Picoband®
Cat # A05900-1

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.