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- Table of Contents
SPR · Western blot design guide
SPR Western Blot Planning Guide
Plan a SPR Western blot around the catalog-observed 28 kDa band, image-backed A00416-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SPR 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 | 28 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Adrenal gland | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 28 kDa |
| ⓘ | Localization | Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse |
Section 1
Real Curated SPR Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A00416-1 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected SPR Western Blot Band Size?
Use the product-observed 28 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 28 kDa | Matches the authoritative product WB observation. |
| 28 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected SPR appearancePlan around 28 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 28 kDa; use this as the primary experimental expectation. |
| Calculated mass | 28 kDa from UniProt P35270; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A00416-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 28 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing SPR band
A00416-1 · SPR
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SPR Western blot
🧪Use Adrenal gland as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Adrenal gland (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for SPR
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Colon | Reported tissue cells | High | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cervix | Reported tissue cells | High | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced SPR Western Blot Tips
Deeper troubleshooting and optimisation questions for SPR, answered from its protein features.
Which band should guide the blot?
Use 28 kDa, the observation attached to the authoritative A00416-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 28 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 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 SPR Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00416-1 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
SPR Western Blot Reagents
Human/Mouse-reactive SPR Western blot reagents with authoritative product imagery.
Real WB data
Anti-SPR Antibody Picoband®
Cat # A00416-1
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
- UniProt P35270
- Human Protein Atlas — SPR
- A00416-1 product record
- PMC4013099 — Novel interaction of ornithine decarboxylase with sepiapterin reductase regulates neuroblastoma cell proliferation (Journal of molecular biology, 2014)
- PMC7539867 — The knockdown of the sepiapterin reductase gene suppresses the proliferation of breast cancer by inducing ROS-mediated apoptosis (International journal of clinical and experimental pathology, 2020)