SRD5A1 · Western blot design guide

SRD5A1 Western Blot Planning Guide

Plan a SRD5A1 Western blot around the catalog-observed 29.5 kDa band, image-backed A03464-2 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 SRD5A1 (SRD5A1): expected band 29.5 kDa, antibody A03464-2, and guide-derived SDS-PAGE protocol steps
SRD5A1 Western blot protocol sheet — expected band 29.5 kDa, antibody A03464-2, controls and PMC citations. Open the full SRD5A1 WB guide →

SRD5A1 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 29.5 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Kidney
Negative control ⓘ Bone marrow
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 29.5 kDa
ⓘ Localization Microsome membrane / Endoplasmic reticulum membrane
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated SRD5A1 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, short transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA03464-2 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 SRD5A1 Western Blot Band Size?

Use the product-observed 29.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
29.5 kDaMatches the authoritative product WB observation.
29.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected SRD5A1 appearancePlan around 29.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band29.5 kDa; use this as the primary experimental expectation.
Calculated mass29.5 kDa from UniProt P18405; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A03464-2.
Why is my band missing or off?
SituationLikely causeNext action
29.5 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 SRD5A1 Western blot

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

HPA tissue expression evidence for SRD5A1

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
Kidney Reported tissue cells High Protein (HPA) HPA →
Fallopian tube Reported tissue cells High Protein (HPA) HPA →
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced SRD5A1 Western Blot Tips

Deeper troubleshooting and optimisation questions for SRD5A1, answered from its protein features.

Which band should guide the blot?
Use 29.5 kDa, the observation attached to the authoritative A03464-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 29.5 kDa expectation.
Which positive control should I start with?
Start with Kidney, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Bone marrow 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 SRD5A1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A03464-2 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

SRD5A1 Western Blot Reagents

Human/Mouse/Rat-reactive SRD5A1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for SRD5A1 using A03464-2; observed band 29.5 kDa
Anti-SRD5A1 Antibody Picoband®
Cat # A03464-2

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 P18405
  2. Human Protein Atlas — SRD5A1
  3. A03464-2 product record
  4. PMC7904855 — Suppression of steroid 5α-reductase type I promotes cellular apoptosis and autophagy via PI3K/Akt/mTOR pathway in multiple myeloma (Cell death & disease, 2021)
  5. PMC7484213 — Steroid 5α-Reductase Type I Induces Cell Viability and Migration via Nuclear Factor-κB/Vascular Endothelial Growth Factor Signaling Pathway in Colorectal Cancer (Frontiers in oncology, 2020)
  6. PMC12752638 — Hepatic KLF9 Deficiency Inhibits Dehydroepiandrosterone (DHEA)-Induced Polycystic Ovary Syndrome via Liver-Ovary Axis (Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2025)