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- Table of Contents
AGRN · Western blot design guide
AGRN Western Blot Planning Guide
Plan an AGRN Western blot around the catalog-observed 217.3 kDa band, image-backed A04649 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
AGRN 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 | 217.3 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 4-12% gradient | |
| Positive control | Kidney | |
| Negative control | Oral mucosa |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 217.3 kDa |
| ⓘ | Localization | Secreted, extracellular space, extracellular matrix / Synapse |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated AGRN Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Kidney |
| Gel % | 4-12% gradient |
| Load | 20-30 µg total protein per lane |
| Transfer | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A04649 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 AGRN Western Blot Band Size?
Use the product-observed 217.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 217.3 kDa | Matches the authoritative product WB observation. |
| 217.3 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 AGRN appearancePlan around 217.3 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 217.3 kDa; use this as the primary experimental expectation. |
| Calculated mass | 217.3 kDa from UniProt O00468; retain as context. |
| Gel selection | 4-12% gradient; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A04649. |
| Situation | Likely cause | Next action |
|---|---|---|
| 217.3 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 AGRN band
A04649 · AGRN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AGRN Western blot
🧪Use Kidney as the first positive-control candidate and Oral mucosa as the HPA Not detected negative candidate.
Positive control: Kidney (High)
Negative control: Oral mucosa (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for AGRN
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 |
|---|---|---|---|---|
| Kidney | Reported tissue cells | High | Protein (HPA) | HPA → |
| Lung | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced AGRN Western Blot Tips
Deeper troubleshooting and optimisation questions for AGRN, answered from its protein features.
Which band should guide the blot?
Use 217.3 kDa, the observation attached to the authoritative A04649 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 217.3 kDa expectation.
Which positive control should I start with?
Start with Kidney, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Oral mucosa as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 4-12% gradient consistently across the quick facts, protocol table, and poster.
What transfer method to use for AGRN Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04649 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
AGRN Western Blot Reagents
Human/Mouse/Rat-reactive AGRN Western blot reagents with authoritative product imagery.
Real WB data
Anti-Agrin/AGRN Picoband® Antibody
Cat # A04649
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 O00468
- Human Protein Atlas — AGRN
- A04649 product record
- PMC10561154 — Agrin is a novel oncogenic protein in thyroid cancer (Oncology letters, 2023)