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AOX1 / Aldehyde oxidase · IHC design guide
Design Immunohistochemistry for AOX1
Plan AOX1 paraffin IHC around cytoplasmic staining in hepatocytes, renal tubules, Leydig cells and adrenal glandular cells (HPA tissue IHC). Use liver hepatocytes as a positive control (HPA tissue IHC) and titrate catalog antibody A02144-1 at 1:100–1:300 (datasheet).
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 | Cytoplasm (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic 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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes lack IHC signal despite reported adipose expression (HPA tissue IHC; UniProt) | |
| Regulation | No significant human sex difference (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–1338 chain (UniProt) |
Section 1
Recommended AOX1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with 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); use each article’s stated retrieval when reproducing its protocol.
Section 2
What Is the Expected AOX1 Staining Pattern?
AOX1 should appear mainly as cytoplasmic staining in hepatocytes, adrenal glandular cells and Leydig cells (HPA tissue IHC: High in each; Enhanced reliability). A cytoplasmic pattern agrees with its reported location and lack of a transmembrane segment (UniProt Q06278: Cytoplasm; no transmembrane segment). HPA’s tissue profile also mentions renal tubules, but its detailed kidney entry rates proximal tubule cell bodies Low (HPA tissue IHC).
| Strong cytoplasmic staining in hepatocytes, adrenal glandular cells or Leydig cells. | This fits the expected cell types and compartment (HPA tissue IHC: High in these cells; cytoplasmic expression). Judge each cell population separately: a strongly stained liver section does not establish that staining in every neighboring cell is specific (general IHC practice). |
| Predominantly nuclear, membrane-like or extracellular staining, with little cytoplasmic signal. | That distribution conflicts with the main expected location (UniProt Q06278: Cytoplasm; HPA tissue IHC: cytoplasmic expression). Check whether counterstain, tissue deposits or the detection system is being mistaken for antibody signal before interpreting an unexpected compartment (general IHC practice). |
| Strong staining in cells listed as undetected, such as adipocytes or appendix glandular cells. | This conflicts with the reported tissue pattern (HPA tissue IHC: Not detected in adipocytes and appendix glandular cells). Cross-reactivity or detection background is possible, but appearance alone cannot identify the cause; compare a no-primary control and an independently validated antibody (general IHC practice). |
| Broad, diffuse chromogen across cells and extracellular spaces, obscuring boundaries. | Such background cannot establish AOX1-positive cells or cytoplasmic localisation (general IHC practice). Compare no-primary and known-positive sections processed together; diffuse signal persisting without primary antibody points toward the detection workflow rather than demonstrated AOX1 staining (general IHC practice). |
| No cytoplasmic signal in a liver section expected to serve as a positive control. | An absent result conflicts with abundant liver expression and High hepatocyte staining (UniProt Q06278: tissue specificity; HPA tissue IHC). First check tissue identity and assay controls, then review the antibody’s documented IHC-P procedure and detection steps (general IHC practice). |
💡Expected AOX1 appearanceCall a result positive when hepatocytes, adrenal glandular cells or Leydig cells show clear cytoplasmic staining at the reported High level (HPA tissue IHC); dominant nuclear, membrane-like or diffuse tissue-wide colour is discordant (UniProt Q06278: Cytoplasm; general IHC practice).
| Cell and tissue selection | Liver hepatocytes, adrenal glandular cells and testicular Leydig cells are reported High; pancreatic exocrine cells and kidney proximal tubule cell bodies are Low (HPA tissue IHC). Use a High population to judge whether the assay can detect AOX1; weak staining in a Low population is less decisive (general IHC practice). |
| Compartment and protein topology | AOX1 is cytoplasmic and has no transmembrane segment or signal peptide (UniProt Q06278). Assess colour within cell bodies rather than treating a membrane rim or extracellular deposit as the expected pattern (general IHC practice). Topology does not establish how fixation or retrieval affects this antibody. |
| Strength of tissue evidence | The tissue profile is rated Enhanced because antibody staining is highly consistent with RNA expression data (HPA tissue IHC). HPA lists HPA040199 and HPA040215 as Enhanced for IHC (HPA antibodies). These ratings support the reported pattern; they do not validate an unspecified antibody or explain an anomalous section. |
| Different evidence for IF/ICC | HPA reports cytosol as the supported main ICC-IF location; nuclear bodies, microtubules and cytokinetic bridge are uncertain additional locations (HPA subcellular). Those uncertain observations should not replace cytoplasmic staining as the IHC interpretation criterion (HPA tissue IHC: cytoplasmic expression). |
| Situation | Likely cause | Next action |
|---|---|---|
| The liver positive control has no convincing hepatocyte staining. | The result disagrees with High hepatocyte staining (HPA tissue IHC); tissue selection or an assay step may need review (general IHC practice). | Verify the section and primary antibody, then check the antibody’s documented IHC-P retrieval, dilution and detection instructions; process a known-positive section alongside the sample (general IHC practice). |
| The whole section is brown, including spaces between cells. | Diffuse colour prevents a cell-specific call; detection background, including endogenous enzyme activity in chromogenic assays, is possible (general IHC practice). | Compare a no-primary section, inspect blocking and detection controls, and score only resolved intracellular staining after background is controlled (general IHC practice). |
| Nuclei or cell borders stain more strongly than cytoplasm. | A dominant nuclear or membrane-like pattern conflicts with AOX1’s reported cytoplasmic location (UniProt Q06278; HPA tissue IHC). | Recheck compartment assignment against the counterstain and morphology; compare the no-primary control and a documented IHC-positive tissue before assigning AOX1 positivity (general IHC practice). |
| Adipocytes or appendix glandular cells stain strongly. | Both cell populations are reported Not detected by tissue IHC (HPA tissue IHC). The stain may reflect cross-reactivity or detection background; the slide alone cannot distinguish them (general IHC practice). | Compare staining in a High positive population, include a no-primary control, and seek confirmation with an independently validated antibody if the unexpected signal persists (general IHC practice). |
| Kidney proximal tubules stain weakly despite the tissue summary mentioning renal tubules. | The detailed kidney entry rates proximal tubule cell bodies Low, while the profile summary names renal tubules among prominent sites (HPA tissue IHC). | Use the detailed cell-level rating when scoring the section; compare a High positive tissue before treating weak kidney staining as assay failure (HPA tissue IHC; general IHC practice). |
| Can the IHC pattern be used as the expected IF/ICC result? | HPA supports cytosol as the main ICC-IF location, while its additional nuclear-body, microtubule and cytokinetic-bridge locations are uncertain (HPA subcellular). | For IF/ICC, assess cytosolic localisation using the separate IF/ICC guide; do not infer an IF protocol or expected signal intensity from tissue IHC ratings (HPA subcellular; HPA tissue IHC). |
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: hepatocytes should show high AOX1 staining (HPA: High in hepatocytes). Use appendix glandular cells as the tissue negative (HPA: Not detected in appendix glandular cells); on the liver slide, assess non-hepatocyte cells as candidate internal negatives, expecting background-level cytoplasmic signal only after confirming their identity and comparing them with the no-primary control (standard IHC practice).
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; an isotype control matched to the primary antibody’s host species, class, and monoclonal or polyclonal format; and an AOX1 knockout or peptide-block control (standard IHC practice; A02144-1 caption: peptide block). For chromogenic liver IHC, quench endogenous peroxidase and block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: AOX1 is cytoplasmic (UniProt Q06278: Cytoplasm), but the supplied evidence reports no target-specific fixation window or fixation effect, and the selected A02144-1 paraffin-section caption does not state a fixative (A02144-1 caption). Antigen-retrieval dependence is unreported; optimize retrieval empirically for paraffin sections (standard IHC practice). HPA provides ICC-IF images in ASC52telo, U-251MG, and U2OS cells (HPA: subcellular), but the supplied evidence does not establish that IF or frozen sections are easier; account for possible liver pigment or endogenous peroxidase background when interpreting chromogenic staining (standard IHC practice).
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 in paraffin sections by checking retrieval, cytoplasmic localisation, tissue controls and background before interpreting chromogenic signal.
How should I retrieve AOX1 in paraffin sections with weak staining?
Start with citrate buffer at pH 6.0 for heat induced epitope retrieval at 95–98 °C for 20 minutes (page retrieval rule). Cool sections consistently, then compare staining with a liver hepatocyte control, where AOX1 expression is high (HPA tissue IHC: liver hepatocytes High). If signal remains weak, test a second retrieval condition on matched serial sections while holding antibody concentration and detection settings constant; this is a troubleshooting comparison, not an established AOX1 requirement (standard IHC practice). Judge improvement by stronger cellular cytoplasmic staining without a parallel rise in diffuse background (UniProt Q06278: Cytoplasm; standard IHC practice).
Does fixation explain weak or uneven AOX1 IHC staining?
AOX1 specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (selected A02144-1 caption: fixative not stated). The catalog image shows staining in paraffin embedded human lung carcinoma tissue, but its caption does not establish a fixation method (selected A02144-1 tissue IHC caption). Record the fixative and processing history for each specimen, then compare similarly processed sections with identical retrieval and detection settings (standard IHC practice). If staining varies across a block, inspect morphology and compare intact interior tissue with damaged or poorly preserved areas before attributing the difference to AOX1 expression (standard IHC practice).
Where should convincing AOX1 staining appear in a section?
Expect predominantly cellular cytoplasmic staining: AOX1 is annotated in the cytoplasm, and its supported subcellular location is cytosol (UniProt Q06278: Cytoplasm; HPA subcellular: Cytosol supported). AOX1 has no transmembrane segment, so a crisp membrane outline alone does not match its recorded topology (UniProt Q06278 topology). Evaluate staining within the expected cells against surrounding tissue; HPA reports abundant cytoplasmic expression in hepatocytes, renal tubules, Leydig cells and adrenal glandular cells (HPA tissue IHC profile). Treat an isolated nuclear or membrane pattern cautiously, while recognizing that HPA lists nuclear bodies only as an uncertain additional location (HPA subcellular: Nuclear bodies uncertain).
Could AOX1 epitope selection explain discordant IHC patterns?
The supplied AOX1 record lists 0 isoforms and one chain spanning residues 1–1338, so a staining difference cannot be assigned to a documented isoform here (UniProt Q06278 processing and isoforms). The protein contains a 2Fe–2S region at residues 5–92 and an FAD binding region at residues 236–421, but no antibody epitope position is supplied (UniProt Q06278 domains; supplied antibody evidence). A phosphoserine at residue 1068 is recorded; its effect on this antibody’s staining is unknown (UniProt Q06278 modified residues; supplied antibody evidence). Compare retrieval conditions and include the catalog image’s peptide blocked control when available, without treating peptide blocking alone as proof of cellular specificity (selected A02144-1 tissue IHC caption; standard IHC practice).
How can IF help assess an ambiguous AOX1 IHC pattern?
Use IF as a separate corroborating experiment: HPA supports a mainly cytosolic AOX1 location, while this page’s catalog evidence is a paraffin section IHC image (HPA subcellular: Cytosol supported; selected A02144-1 tissue IHC caption). Multiplex AOX1 with a validated marker for the expected cell population, such as hepatocytes in liver, and check that signals occupy the appropriate cells rather than merely overlap in crowded tissue (HPA tissue IHC: liver hepatocytes High; standard IF practice). Choose fluorophores after measuring tissue autofluorescence and include single stain controls for spectral separation (standard IF practice). Because AOX1 lacks a transmembrane segment and is cytosolic, permeabilise fixed cells sufficiently for intracellular antibody access, then verify morphology and background (UniProt Q06278 topology; HPA subcellular: Cytosol supported; standard IF practice).
How do I reduce diffuse or false positive AOX1 chromogenic staining?
First compare a no primary control and an appropriate low expression compartment with the stained section to locate background from detection reagents or tissue (standard IHC practice). HPA reports AOX1 as not detected in bronchial respiratory epithelial cells and appendix glandular cells, although those observations do not guarantee a blank result under every assay condition (HPA tissue IHC). For peroxidase based chromogenic detection, include an endogenous peroxidase block and inspect the no primary control before interpreting brown signal as AOX1 (standard IHC practice). Titrate the primary antibody and detection exposure on matched sections, preserving hepatocyte cytoplasmic staining as a positive reference (HPA tissue IHC: liver hepatocytes High; standard IHC practice).
How should I score AOX1 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring because the supported AOX1 location is cytosolic (HPA subcellular: Cytosol supported). For comparable sections, report an H-score using the percentage of cells at each intensity, or report the percentage of cytoplasm positive cells with the same threshold throughout (standard IHC practice). If distribution matters, report positive cell density per mm² of viable tissue as a separate measure (standard IHC practice). Normalise counts to the number of evaluable target cells or viable tissue area, and compare batches against a shared control section with fixed imaging and scoring settings (standard IHC practice).
When is an unexpected AOX1 IHC signal likely to be artefactual?
Give greatest weight to reproducible cytoplasmic signal in biologically plausible cells, such as hepatocytes, which HPA scores High for AOX1 (UniProt Q06278: Cytoplasm; HPA tissue IHC: liver hepatocytes High). A membrane only pattern, strong signal in necrotic tissue, or staining confined to section edges warrants review of morphology, retrieval and detection controls (UniProt Q06278 topology; standard IHC practice). Check a no primary section and the endogenous peroxidase block when brown precipitate appears outside intact cells (standard IHC practice). The selected catalog image includes a peptide blocked companion image, which supports an antibody competition comparison but does not by itself establish that every stained cell contains AOX1 (selected A02144-1 tissue IHC caption; standard IHC practice).
Boster reagents
Best AOX1 / Aldehyde oxidase IHC Antibodies
Two anti-AOX1 antibodies list IHC use (catalog: applications). A02144-1 shows human lung carcinoma paraffin-section IHC and lists IF use with human and rat reactivity (catalog: A02144-1 IHC image caption/applications/reactivity).
Real IHC data
Anti-AOX1/Aldehyde Oxidase Antibody
Cat # A02144-1
The rendered A02144-1 card shows IHC on paraffin-embedded human lung carcinoma, with peptide blocking shown alongside the stain (catalog: A02144-1 IHC image caption). A02144-1 also lists IF and human/rat reactivity, but supplies no IF image (catalog: A02144-1 applications/reactivity/IF image alts).
Which to pick: For human paraffin-section tissue IHC, choose A02144-1 based on its own image caption; the fixative is unreported (catalog: A02144-1 IHC image caption). For IF, A02144-1 lists human/rat reactivity and a 1:50 dilution; ICC validation and IF imagery are unreported (catalog: A02144-1 applications/reactivity/IF dilution/IF image alts). For mouse tissue IHC, consider A02144-2, a rabbit polyclonal with listed IHC use and human/mouse/rat reactivity; it supplies no IHC image or processing caption (catalog: A02144-2 host/dilution_raw/applications/reactivity/IHC image alts).
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 Cytoplasm (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 paraffin sections (standard IHC practice; not target-specific); Caveat: Adipocytes lack IHC signal despite reported adipose expression (HPA tissue IHC; UniProt); Isoform / epitope: No annotated isoforms; one 1–1338 chain (UniProt); Regulation: No significant human sex difference (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.