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- Table of Contents
AOX1 / Aldehyde oxidase · Western blot design guide
Design a Western Blot for AOX1
Real validated AOX1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AOX1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
AOX1 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 | ~147.9 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +2 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated AOX1 Western Blot Protocols
The A02144-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF-7, sp2/0, PC12 (catalog A02144-2) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02144-2; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected AOX1 Western Blot Band Size?
AOX1 has a predicted 147.9 kDa monomer; homodimerization could affect migration, but no empirical band size or demonstrated shift is supplied.
| Band near 148 kDa | Consistent with the predicted AOX1 monomer; confirm identity with antibody controls |
| Band near 296 kDa | Could reflect retained AOX1 homodimer |
| Band in cytoplasmic lysate | Consistent with AOX1 cytoplasmic localization |
| No visible shift after phosphatase treatment | Ser1068 phosphorylation need not cause a detectable mobility change |
💡Expected AOX1 appearanceUniProt predicts a 147.9 kDa AOX1 monomer, but no empirical band size is supplied; use antibody specificity and AOX1 depletion controls to establish band identity.
| Predicted monomer mass | Places the expected monomer near 147.9 kDa |
| Sequence-derived mass | Gives the same monomer prediction as 147918 Da |
| AOX1 homodimer | Could yield a band near twice the monomer mass if the complex remains intact |
| Homodimer dissociation | Would favor monomer migration near the predicted mass |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | AOX1 may be scarce in the sampled cells | Check AOX1 expression and include a verified positive lysate |
| Band higher than expected | An AOX1 homodimer may remain intact | Compare denaturing conditions and verify the band by AOX1 depletion |
| Band lower than expected | Band identity or protein integrity is uncertain | Check molecular weight markers and test whether AOX1 depletion removes the band |
| Multiple bands | Monomer and retained homodimer are possible | Compare denaturing conditions and confirm each band with AOX1 depletion |
| Weak or no signal | AOX1 abundance or detection sensitivity may be insufficient | Verify loading and use a confirmed AOX1 positive control |
Experiment evidence
Western blot images showing AOX1 band
A02144-2 · AOX1
A02144-1 · AOX1
A02144-1 · AOX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AOX1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AOX1 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cytoplasmic AOX1 should be accessible in tissue lysates, with HPA indicating clear positive and negative tissue choices.
HPA tissue expression evidence for AOX1
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.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Liver | hepatocytes | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AOX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for AOX1, answered from its protein features.
How should AOX1 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could AOX1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence, so it does not support assigning multiple bands to AOX1 isoforms. Check any extra bands for antibody specificity before interpreting them.
Which AOX1 modification is relevant to phospho-sensitive detection?
PTM · UniProt lists phosphoserine at position 1068. Use UniProt numbering when comparing this site with an antibody or paper, whose numbering may differ. The listed site alone does not establish a detectable band shift.
Does this guide establish induction of AOX1?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for AOX1 Western blot?
Transfer · AOX1 is a 147.9 kDa protein. Choose transfer conditions that recover proteins near 148 kDa, and assess transfer with a total-protein stain or the post-transfer gel. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02144-2 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should AOX1 bands be quantified?
Quantitation · Quantify a validated AOX1 band within the assay’s linear signal range and normalize to a suitable total-protein measure. AOX1 is listed as cytoplasmic, so keep the sample fraction consistent across comparisons. Do not treat band intensity alone as a measure of enzyme activity.
Why might an AOX1 band differ from its predicted mass?
Interpretation · AOX1 has a predicted mass of 147.9 kDa, but no observed band size is supplied. Its listed phosphoserine at UniProt position 1068 does not by itself establish a visible shift or explain a mass difference. Check the band against a molecular weight marker.
AOX1 is listed as a homodimer, making dimerization a possibility when interpreting high-mass species. The feature does not show that a dimer survives the chosen sample preparation or appears on a Western blot. Verify band identity before assigning it to AOX1.
AOX1 is listed as a homodimer, making dimerization a possibility when interpreting high-mass species. The feature does not show that a dimer survives the chosen sample preparation or appears on a Western blot. Verify band identity before assigning it to AOX1.
Boster reagents
AOX1 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Aldehyde oxidase AOX1 Antibody
Cat # A02144-2
Real WB data
Anti-AOX1/Aldehyde Oxidase Antibody
Cat # A02144-1
Two the supplier anti-AOX1 antibodies are listed with WB images. A02144-2 has a caption naming MCF-7, sp2/0, and PC12 lysates at 1:500; A02144-1 reports various cells at 1:2000 without naming them. No publication evidence is supplied.
Which to pick: Choose A02144-2 for reported human, mouse, or rat reactivity and the named WB lysate examples. A02144-1 lists human and rat reactivity and has a WB image, but its caption does not identify the cells tested.
Source: BosterBio AOX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q06278.
- Human Protein Atlas. AOX1 tissue expression.
- PMC13304650 — target-verified WB comparison
- PMC12891850 — target-verified WB comparison
- PMC8058741 — target-verified WB comparison