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- Table of Contents
ASPH · Western blot design guide
ASPH Western Blot Planning Guide
Plan an ASPH Western blot around the catalog-observed 85.9 kDa band, image-backed PB9478 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
ASPH 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 | 85.9 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Adipose tissue | |
| Negative control | Heart muscle |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 85.9 kDa |
| ⓘ | Localization | Endoplasmic reticulum membrane / Sarcoplasmic reticulum membrane |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated ASPH Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adipose tissue |
| Gel % | 8-10% |
| 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 | PB9478 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 ASPH Western Blot Band Size?
Use the product-observed 85.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 85.9 kDa | Matches the authoritative product WB observation. |
| 85.9 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 ASPH appearancePlan around 85.9 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 85.9 kDa; use this as the primary experimental expectation. |
| Calculated mass | 85.9 kDa from UniProt Q12797; retain as context. |
| Gel selection | 8-10%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with PB9478. |
| Situation | Likely cause | Next action |
|---|---|---|
| 85.9 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 ASPH band
PB9478 · ASPH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ASPH Western blot
🧪Use Adipose tissue as the first positive-control candidate and Heart muscle as the HPA Not detected negative candidate.
Positive control: Adipose tissue (High)
Negative control: Heart muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for ASPH
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 |
|---|---|---|---|---|
| Adipose tissue | Reported tissue cells | High | Protein (HPA) | HPA → |
| Adrenal gland | Reported tissue cells | High | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | High | Protein (HPA) | HPA → |
| Caudate | Reported tissue cells | High | Protein (HPA) | HPA → |
Section 3
Advanced ASPH Western Blot Tips
Deeper troubleshooting and optimisation questions for ASPH, answered from its protein features.
Which band should guide the blot?
Use 85.9 kDa, the observation attached to the authoritative PB9478 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 85.9 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, 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 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for ASPH Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9478 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
ASPH Western Blot Reagents
Human/Mouse/Rat-reactive ASPH Western blot reagents with authoritative product imagery.
Real WB data
Anti-Aspartate beta hydroxylase/ASPH Antibody Picoband®
Cat # PB9478
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 Q12797
- Human Protein Atlas — ASPH
- PB9478 product record
- PMC5541716 — Aspartate β-hydroxylase disrupts mitochondrial DNA stability and function in hepatocellular carcinoma (Oncogenesis, 2017)
- PMC5934571 — MiR-200a inhibits cell proliferation and EMT by down-regulating the ASPH expression levels and affecting ERK and PI3K/Akt pathways in human hepatoma cells (American journal of translational research, 2018)
- PMC5878085 — Phosphorylation Modulates Aspartyl-(Asparaginyl)-β Hydroxylase Protein Expression, Catalytic Activity and Migration in Human Immature Neuronal Cerebellar Cells (Cell biology : research & therapy, 2013)
