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- Table of Contents
DEFA1 · Western blot design guide
DEFA1 Western Blot Planning Guide
Plan a DEFA1 Western blot around the catalog-observed 10.2 kDa band, image-backed A10546 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
DEFA1 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 | 10.2 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 15% | |
| Positive control | Spleen | |
| Negative control | Appendix |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 10.2 kDa |
| ⓘ | Localization | Secreted |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Rat |
Section 1
Real Curated DEFA1 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Bone marrow |
| Gel % | 15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.2 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A10546 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 DEFA1 Western Blot Band Size?
Use the product-observed 10.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 10.2 kDa | Matches the authoritative product WB observation. |
| 10.2 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 DEFA1 appearancePlan around 10.2 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 10.2 kDa; use this as the primary experimental expectation. |
| Calculated mass | 10.2 kDa from UniProt P59665; retain as context. |
| Gel selection | 15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A10546. |
| Situation | Likely cause | Next action |
|---|---|---|
| 10.2 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 DEFA1 band
A10546 · DEFA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DEFA1 Western blot
🧪Use Spleen as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.
Positive control: Spleen (High)
Negative control: Appendix (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for DEFA1
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.
Section 3
Advanced DEFA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DEFA1, answered from its protein features.
Which band should guide the blot?
Use 10.2 kDa, the observation attached to the authoritative A10546 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 10.2 kDa expectation.
Which positive control should I start with?
Start with Spleen, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Appendix as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for DEFA1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A10546 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
DEFA1 Western Blot Reagents
Human/Rat-reactive DEFA1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-Alpha Defensin 1/DEFA1 Antibody Picoband®
Cat # A10546
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 P59665
- Human Protein Atlas — DEFA1
- A10546 product record
- PMC12652293 — Defensin-Rich Platelets Drive Pro-Tumorigenic Programs in Pancreatic Adenocarcinoma (International journal of molecular sciences, 2025)
- PMC3069341 — An informatics-assisted label-free approach for personalized tissue membrane proteomics: case study on colorectal cancer (Molecular & cellular proteomics : MCP, 2011)
- PMC11880931 — DEFA1, Primarily Expressed at the Invasive Tumor Front, Promotes OSCC Cell Invasion and Tumor Growth (Cancer genomics & proteomics, 2025)