ALAS1 · Western blot design guide

ALAS1 Western Blot Planning Guide

Plan an ALAS1 Western blot around the catalog-observed 70.6 kDa band, image-backed A10405-1 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for ALAS1 (ALAS1): expected band 70.6 kDa, antibody A10405-1, and guide-derived SDS-PAGE protocol steps
ALAS1 Western blot protocol sheet — expected band 70.6 kDa, antibody A10405-1, controls and PMC citations. Open the full ALAS1 WB guide →

ALAS1 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 70.6 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 70.6 kDa
ⓘ Localization Mitochondrion inner membrane
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated ALAS1 Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysateAdipose tissue
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA10405-1 at datasheet starting dilution
Primary incubationOvernight at 4 °C with gentle agitation
SecondarySpecies-matched HRP conjugate at validated dilution
Wash3 × 5 min in TBST
DetectionChemiluminescent substrate
ExposureBracket exposures to avoid saturation
Section 2

What Is the Expected ALAS1 Western Blot Band Size?

Use the product-observed 70.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
70.6 kDaMatches the authoritative product WB observation.
70.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ALAS1 appearancePlan around 70.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band70.6 kDa; use this as the primary experimental expectation.
Calculated mass70.6 kDa from UniProt P13196; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A10405-1.
Why is my band missing or off?
SituationLikely causeNext action
70.6 kDaMatches the authoritative product WB observation.Confirm with orthogonal controls and the linked product record.
Additional bandMay reflect processing, modification, or non-specific signal.Run a dilution series and compare positive/negative controls.
Weak signalTarget abundance or transfer may be limiting.Verify transfer, increase positive-control abundance, and bracket exposure.

Sample controls for ALAS1 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 ALAS1

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

TissueCell typeLevelEvidenceSource
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Cerebellum Reported tissue cells High Protein (HPA) HPA →
Adipose tissue Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Section 3

Advanced ALAS1 Western Blot Tips

Deeper troubleshooting and optimisation questions for ALAS1, answered from its protein features.

Which band should guide the blot?
Use 70.6 kDa, the observation attached to the authoritative A10405-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 70.6 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 ALAS1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A10405-1 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

ALAS1 Western Blot Reagents

Human/Mouse/Rat-reactive ALAS1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for ALAS1 using A10405-1; observed band 70.6 kDa
Anti-Alas1 Antibody Picoband®
Cat # A10405-1

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

  1. UniProt P13196
  2. Human Protein Atlas — ALAS1
  3. A10405-1 product record
  4. PMC6804493 — Synergistic antitumor activity of artesunate and HDAC inhibitors through elevating heme synthesis via synergistic upregulation of ALAS1 expression (Acta pharmaceutica Sinica. B, 2019)
  5. PMC12195290 — Comparison of Pyrazinamide with Isoniazid for Their Effects on the Heme Biosynthetic Pathway in Mouse Liver (Metabolites, 2025)