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- Table of Contents
ARFGAP1 · Western blot design guide
ARFGAP1 Western Blot Planning Guide
Plan an ARFGAP1 Western blot around the catalog-observed 44.7 kDa band, image-backed A04959 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
ARFGAP1 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 | 44.7 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Cervix | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 44.7 kDa |
| ⓘ | Localization | Cytoplasm / Golgi apparatus |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated ARFGAP1 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A04959 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 ARFGAP1 Western Blot Band Size?
Use the product-observed 44.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 44.7 kDa | Matches the authoritative product WB observation. |
| 44.7 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 ARFGAP1 appearancePlan around 44.7 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 44.7 kDa; use this as the primary experimental expectation. |
| Calculated mass | 44.7 kDa from UniProt Q8N6T3; 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 A04959. |
| Situation | Likely cause | Next action |
|---|---|---|
| 44.7 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 ARFGAP1 band
A04959 · ARFGAP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ARFGAP1 Western blot
🧪Use Cervix as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Cervix (Medium)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for ARFGAP1
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 |
|---|---|---|---|---|
| Cervix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Cerebral cortex | Reported tissue cells | Medium | Protein (HPA) | HPA → |
Section 3
Advanced ARFGAP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ARFGAP1, answered from its protein features.
Which band should guide the blot?
Use 44.7 kDa, the observation attached to the authoritative A04959 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 44.7 kDa expectation.
Which positive control should I start with?
Start with Cervix, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Heart muscle 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 ARFGAP1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04959 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
ARFGAP1 Western Blot Reagents
Human/Mouse/Rat-reactive ARFGAP1 Western blot reagents with authoritative product imagery.
Real WB data
Anti-ARFGAP1 Antibody Picoband®
Cat # A04959
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 Q8N6T3
- Human Protein Atlas — ARFGAP1
- A04959 product record
- PMC13139335 — LRRK2 regulates ArfGAP1 membrane localization, activity and neuronal integrity via phosphorylation within its lipid-sensing ALPS2 motif (Frontiers in molecular neuroscience, 2026)
- PMC8204869 — ArfGAP1 inhibits mTORC1 lysosomal localization and activation (The EMBO journal, 2021)
- PMC3087831 — ARFGAP1 promotes AP-2-dependent endocytosis (Nature cell biology, 2011)
