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AOX1 / Aldehyde oxidase · IHC design guide
Design Immunohistochemistry for AOX1
Plan AOX1 staining in paraffin sections using its cytoplasmic tissue pattern and an IHC-validated antibody (HPA tissue IHC; datasheet). Compare positive hepatocytes with an appropriate negative control, and score staining by cell type (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AOX1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasm in hepatocytes, renal tubules, Leydig and adrenal cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+2 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes are unstained despite adipose tissue expression (HPA tissue IHC; UniProt) | |
| Regulation | No significant human sex effect (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one cytoplasmic chain (UniProt) |
Section 1
Recommended AOX1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published AOX1 chromogenic IHC protocols (PMC3415238; PMC8287305; PMC13304650; PMC8058741).
| Sample | Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A02144-1) |
| Fixation | Image fixative and duration unreported (datasheet A02144-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-AOX1, 1:100-1:300 (datasheet A02144-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AOX1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression most abundant in hepatocytes, renal tubules, Leydig cells and adrenal glandular cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the ccRCC study used Tris-EDTA pH 9.0 (PMC8287305: IHC).
Section 2
What Is the Expected AOX1 Staining Pattern?
AOX1 should show cytoplasmic staining in hepatocytes, adrenal glandular cells and Leydig cells, where HPA reports high staining (HPA tissue IHC). Its reported location is cytoplasmic, consistent with a protein lacking a transmembrane segment (UniProt Q06278). HPA rates the tissue IHC pattern “Enhanced,” citing high consistency between antibody staining and RNA expression data (HPA tissue IHC).
| Strong cytoplasmic staining in hepatocytes, adrenal glandular cells or Leydig cells. | This fits the reported high staining in those cell populations (HPA tissue IHC) and AOX1’s cytoplasmic location (UniProt Q06278). Judge the signal in the named cells, since staining across an entire tissue section is less informative than its cellular distribution (general IHC practice). |
| Predominantly nuclear or sharply membrane-bound staining. | These compartments depart from the expected cytoplasmic IHC pattern (HPA tissue IHC; UniProt Q06278 topology). Review morphology and controls before interpreting such signal as AOX1; compartment mismatch raises concern for nonspecific staining or an imaging or scoring artefact (general IHC practice). |
| Strong staining in adipocytes or bronchial respiratory epithelial cells. | HPA reports AOX1 as not detected in these specific cell types (HPA tissue IHC). Check whether the stained cells were identified correctly, then assess background and detection controls; unexpected signal may reflect cross-reactivity or endogenous detection activity (general IHC practice). |
| Diffuse colour across cells and tissue spaces, with little cellular distinction. | A uniform haze does not reproduce the cell-restricted cytoplasmic pattern reported for AOX1 (HPA tissue IHC). Compare with a no-primary control and inspect washing, blocking and detection conditions for background that could obscure genuine staining (general IHC practice). |
| No convincing signal in hepatocytes in a liver section. | This conflicts with HPA’s high hepatocyte staining and UniProt’s report of abundant liver expression (HPA tissue IHC; UniProt Q06278). Confirm that tissue and cells are suitable, then check the antibody’s IHC-P instructions, retrieval, detection reagents and run controls (general IHC practice). |
💡Expected AOX1 appearanceCall a section positive when the named cells show clear cytoplasmic staining, strongest in hepatocytes, adrenal glandular cells or Leydig cells (HPA tissue IHC; UniProt Q06278); dominant nuclear, membrane-bound or diffuse tissue-wide colour warrants control review (general IHC practice).
| Which tissue and cells provide a positive reference? | Liver hepatocytes, adrenal glandular cells and testicular Leydig cells are reported as high (HPA tissue IHC). HPA also describes cytoplasmic expression in renal tubules, while its listed proximal-tubule cell-body level is low; use that qualified record when judging kidney (HPA tissue IHC). |
| Can a whole tissue be treated as uniformly negative? | Interpret the named cell type: HPA reports no detection in adipocytes from adipose tissue or breast, among other listed cells (HPA tissue IHC). UniProt reports expression in adipose tissue without specifying those cells (UniProt Q06278); these records do not establish that every adipose-tissue cell is negative. |
| How strong is the tissue-pattern evidence? | HPA assigns tissue IHC “Enhanced” reliability and lists two rabbit polyclonal antibodies with Enhanced IHC status, HPA040199 and HPA040215 (HPA tissue IHC; HPA antibodies). That supports the reported pattern, while an individual staining run still needs appropriate controls (general IHC practice). |
| Does AOX1 processing suggest a separate secreted staining pattern? | UniProt describes one chain spanning residues 1–1338, no signal peptide or propeptide, and no transmembrane segment (UniProt Q06278). Its reported cytoplasmic location supports cytoplasmic interpretation; these annotations alone do not predict retrieval requirements or fixation sensitivity (UniProt Q06278). |
| What should IF/ICC show? | Mainly cytosolic signal is supported by HPA’s ICC-IF record; nuclear bodies, microtubules and the cytokinetic bridge are additional uncertain locations (HPA subcellular). Treat those additional patterns cautiously. HPA lists HPA040199 as ICC Supported; this IHC section supplies no IF/ICC protocol (HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver positive control is blank or very faint. | High hepatocyte staining is expected, so a blank result may indicate a failed staining run or unsuitable specimen (HPA tissue IHC; general IHC practice). | Verify hepatocytes are present; review the catalog antibody’s IHC-P instructions, retrieval conditions, dilution, detection and run controls (general IHC practice). No AOX1-specific fixation sensitivity is established by the supplied sources. |
| Nuclei dominate the signal. | Dominant nuclear staining conflicts with AOX1’s cytoplasmic location and the reported tissue IHC pattern (UniProt Q06278; HPA tissue IHC). | Recheck compartment boundaries with the counterstain, compare the no-primary control and review antibody and detection conditions before scoring the nuclear signal as specific (general IHC practice). |
| Adipocytes or bronchial respiratory epithelium stain strongly. | HPA reports AOX1 as not detected in those cell types; the apparent signal may reflect cell-identification error, cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). | Confirm cell identity on the section and compare negative and no-primary controls; inspect the detection system for background before assigning AOX1 positivity (general IHC practice). |
| Brown colour spreads beyond cell boundaries. | Diffuse colour lacks the cellular and cytoplasmic definition expected from HPA’s AOX1 pattern (HPA tissue IHC). Background from detection or incomplete washing is a general possibility (general IHC practice). | Compare a no-primary control, inspect tissue morphology and review blocking, washes and detection development using standard IHC controls (general IHC practice). |
| Kidney looks weaker than liver. | The HPA profile mentions renal tubules, but its listed proximal-tubule cell-body level is low, whereas hepatocytes are high (HPA tissue IHC). | Score each identified cell population against its own reported level; do not require kidney intensity to equal liver intensity (HPA tissue IHC). |
| IF/ICC shows nuclear-body or microtubule signal. | HPA labels these additional locations uncertain, while its main cytosolic location is supported (HPA subcellular). | Prioritise cytosolic localisation when interpreting AOX1 and verify unusual structures with suitable imaging controls (HPA subcellular; general IF practice). Consult the separate IF/ICC guide for that application. |
Experiment evidence
IHC & IF images showing AOX1 staining
A02144-1 · IHCImmunohistochemistry analysis of paraffin-embedded human lung carcinoma tissue, using AOX1 Antibody. The picture on the right is blocked wit
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138356-AOX1/tissue/cerebral+cortex#imid_20582219
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000138356-AOX1/tissue/adrenal+gland#imid_20582195
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138356-AOX1/tissue/colon#imid_20582129
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138356-AOX1/tissue/liver#imid_20582130
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA040199; green = AOX1, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA040199; green = AOX1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AOX1 IHC & IF
🧪Run liver first and expect hepatocytes to stain (HPA: High in liver hepatocytes). Use appendix glandular cells as a negative tissue (HPA: Not detected in appendix glandular cells); on the liver slide, assess nonhepatocyte areas for background without assuming every cell there is AOX1-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AOX1 in ASC52telo, U-251MG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; use host-matched nonimmune IgG, with the same isotype if the primary is monoclonal; and compare with peptide-blocked staining as a biological specificity control (selected A02144-1 tissue-IHC caption: peptide block). For liver chromogenic IHC, block endogenous peroxidase and assess tissue pigment when interpreting signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative is unreported in the selected A02144-1 paraffin-section caption. AOX1-specific antigen-retrieval dependence is unreported, so optimize retrieval empirically for the catalog antibody; the supplied evidence does not establish that frozen sections or IF are easier. In liver, endogenous peroxidase and pigment can complicate chromogenic interpretation (standard IHC practice); expected staining is cytoplasmic (UniProt Q06278: Cytoplasm).
HPA tissue IHC evidence for AOX1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended 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 IHC Tips
Troubleshoot AOX1 staining by checking retrieval, cytoplasmic localisation, tissue context and control behaviour in paraffin section IHC.
How should I retrieve AOX1 in paraffin sections if staining is weak?
Begin with citrate pH 6.0 heat induced retrieval at 95–98 °C for 20 min on paraffin sections (page retrieval rule). Keep section thickness, heating vessel and cooling period consistent across the comparison, because retrieval variation can change chromogenic signal (standard IHC practice). If staining remains weak, compare a modest increase in retrieval time with the starting condition while checking tissue integrity (standard IHC practice). Assess the result in hepatocytes or adrenal glandular cells, where AOX1 protein staining is high (HPA tissue IHC). Score cytoplasmic signal rather than nuclear colour as the expected endpoint (UniProt Q06278 localisation; HPA tissue IHC).
Can I attribute weak AOX1 staining to the tissue fixative?
Target specific AOX1 sensitivity to fixation is unknown from the supplied evidence; the selected image identifies paraffin embedded lung carcinoma tissue but reports no fixative (caption A02144-1). Record the actual fixative and fixation duration for each specimen before comparing staining, since these variables can affect IHC antigen accessibility (standard IHC practice). Run sections from the same block through the citrate pH 6.0, 20 min retrieval condition when investigating variability (page retrieval rule). Include a known positive tissue section in the same staining run, such as liver with hepatocyte staining (HPA tissue IHC; standard IHC practice). Do not assign a fixation effect from localisation or tissue distribution alone (UniProt Q06278 localisation; HPA tissue IHC).
Where should convincing AOX1 staining appear in a tissue section?
Expect predominantly cytoplasmic staining in AOX1 positive cells, consistent with its cytoplasmic annotation and supported cytosolic localisation (UniProt Q06278 localisation; HPA subcellular). AOX1 has no transmembrane segment, so a crisp membrane rim without cytoplasmic signal warrants scrutiny (UniProt Q06278 topology). Compare compartment patterns within morphologically intact cells against a positive tissue control, preferably hepatocytes or adrenal glandular cells (HPA tissue IHC; standard IHC practice). Counterstain lightly enough to distinguish cytoplasm from nuclei and inspect both high and low signal areas at the same magnification (standard IHC practice). Treat isolated nuclear staining cautiously: reported nuclear body localisation is uncertain (HPA subcellular).
Could AOX1 isoforms or epitope masking explain discordant staining?
The supplied AOX1 record lists 0 isoforms and a single 1–1338 chain, so an isoform switch is unsupported as an explanation (UniProt Q06278 processing and isoforms). AOX1 has no signal peptide or transmembrane segment, which supports evaluating antibody staining as a cytoplasmic pattern (UniProt Q06278 processing, topology and localisation). Compare adjacent sections with the specified citrate pH 6.0 retrieval and identical detection settings before attributing a weak result to epitope accessibility (page retrieval rule; standard IHC practice). The catalog image includes peptide blocked staining in paraffin embedded lung carcinoma tissue, offering an antibody competition comparison for that image (caption A02144-1). Competition can support binding specificity but does not establish that every stained cell expresses AOX1 (standard IHC interpretation).
How can companion IF help assess an ambiguous AOX1 IHC pattern?
Use IF as a companion localisation check while judging the primary chromogenic result by its tissue morphology and controls (standard IHC practice). Multiplex AOX1 with a validated marker for the expected cell type, such as a hepatocyte marker in liver, because hepatocytes show high AOX1 staining (HPA tissue IHC; standard IF practice). Choose a far red fluorophore, around 650 nm emission, when shorter wavelength tissue autofluorescence obscures signal, and include single channel controls (standard IF practice). For an intracellular AOX1 epitope, use a controlled permeabilisation step, such as 0.1% detergent, and compare it with an untreated section (UniProt Q06278 topology and localisation; standard IF practice). AOX1 is mainly cytosolic in the supplied IF localisation evidence (HPA subcellular).
What should I check when AOX1 DAB staining looks diffuse?
First inspect the no primary control and an adjacent positive tissue section stained in the same run; widespread colour in the control points to the detection workflow (standard IHC practice). For peroxidase based DAB detection, verify the peroxide block and wash steps, since endogenous enzyme activity can produce colour without specific antibody binding (standard IHC practice). Reassess blocking, antibody concentration and incubation time if background persists, changing 1 variable per comparison (standard IHC practice). Keep the counterstain light enough to recognise cytoplasmic signal in intact cells (UniProt Q06278 localisation; standard IHC practice). The peptide blocked catalog image offers a comparison for its paraffin embedded lung carcinoma specimen, with fixation unspecified (caption A02144-1).
How should I score AOX1 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because AOX1 is mainly cytosolic and its abundance varies by tissue cell type (HPA subcellular; HPA tissue IHC). An H-score can combine the percentage of cells at intensity levels 0–3, yielding a 0–300 scale; report the scoring rule (standard IHC practice). Alternatively report percentage positive cells or positive cell density per mm² when cell counts are more reproducible than intensity (standard IHC practice). Normalise to the number or area of viable cells of the same annotated type, rather than total section area when tissue composition differs (standard IHC practice). Apply the same threshold, imaging settings and positive control across batches (standard IHC practice).
How do I distinguish true AOX1 staining from tissue artefact?
A credible positive pattern is cytoplasmic within morphologically intact cells of a type expected to express AOX1, such as hepatocytes or adrenal glandular cells (UniProt Q06278 localisation; HPA tissue IHC). Predominantly membrane rim or nuclear staining requires extra scrutiny because AOX1 lacks a transmembrane segment and nuclear body localisation is uncertain (UniProt Q06278 topology; HPA subcellular). Compare central tissue with section edges and exclude necrotic regions from scoring, where uneven staining can distort interpretation (standard IHC practice). Check no primary and peroxidase blocked controls when DAB colour appears in unexpected cells or compartments (standard IHC practice). Interpret the peptide blocked lung carcinoma image only as antibody competition evidence for that specimen (caption A02144-1).
Boster reagents
Best AOX1 / Aldehyde oxidase IHC Antibodies
The catalog includes human-reactive AOX1 antibodies for IHC and IF (catalog: A02144-1, A02144-2); one has a paraffin-section IHC image from human lung carcinoma (A02144-1 image caption).
Real IHC data
Anti-AOX1/Aldehyde Oxidase Antibody
Cat # A02144-1
The rendered card is A02144-1: its IHC image shows paraffin-embedded human lung carcinoma tissue with a peptide-blocked comparison (A02144-1 image caption). IF is listed as an application for A02144-1, with human and rat reactivity, but no IF image is supplied (catalog: A02144-1).
Which to pick: Choose A02144-1 for paraffin-section IHC when a matching image matters; its caption does not report the fixative (A02144-1 image caption). For IF/ICC, A02144-1 is the only listed IF option, though the catalog supplies no IF image or ICC validation (catalog: A02144-1, A02144-2). For mouse tissue IHC, consider A02144-2 because it lists mouse reactivity and IHC, while noting that no IHC image is supplied for that SKU (catalog: A02144-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q06278 (AOXA_HUMAN, Aldehyde oxidase).
- Human Protein Atlas. AOX1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AOX1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nuclear bodies, microtubules and cytokinetic bridge..
- Human Protein Atlas. AOX1 antibody validation summary (2 antibodies).
- Decreased skin-mediated detoxification contributes to oxidative stress and insulin resistance. Experimental diabetes research 2012 — PMC3415238.
- Expression of AOX1 Predicts Prognosis of Clear Cell Renal Cell Carcinoma. Frontiers in genetics 2021 — PMC8287305.
- Connective tissue profiling in keratinized and non-keratinized oral mucosa reveals distinct extracellular and intracellular features. Frontiers in cell and developmental biology 2026 — PMC13304650.
- Epigenetic loss of AOX1 expression via EZH2 leads to metabolic deregulations and promotes bladder cancer progression. Oncogene 2020 — PMC8058741.
- PubMed PMID:8248161 — UniProt-cited evidence.
- PubMed PMID:27406959 — UniProt-cited evidence.
- PubMed PMID:11302742 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AOX1 (Aldehyde oxidase). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02144-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A02144-1); 2. Fixation: Image fixative and duration unreported (datasheet A02144-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-AOX1, 1:100-1:300 (datasheet A02144-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Adipocytes are unstained despite adipose tissue expression (HPA tissue IHC; UniProt); Isoform / epitope: 0 annotated isoforms; one cytoplasmic chain (UniProt); Regulation: No significant human sex effect (UniProt). Evidence: UniProt Q06278, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A02144-1, PMC3415238, PMC8287305. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AOX1/immunohistochemistry-ihc-guide.