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ACOX1 / Peroxisomal acyl-coenzyme A oxidase 1 · IHC design guide
Design Immunohistochemistry for ACOX1
Use liver hepatocytes and intestinal glandular cells as high-staining references for ACOX1 paraffin IHC (HPA tissue IHC). The catalog antibody M03054 is tested for IHC-P at 1:10–1:50 (datasheet: M03054); assess granular cytoplasmic staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACOX1 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 | Granular cytoplasm (HPA tissue IHC); peroxisome (UniProt) | |
| Staining pattern | Hepatocytes and glandular cells show granular cytoplasm (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 | Staining has low concordance with tissue RNA (HPA tissue IHC) | |
| Regulation | Liver RNA is tissue enhanced (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; B/C processing may alter epitope detection (UniProt) |
Section 1
Recommended ACOX1 IHC & IF Protocols
The catalog antibody protocol and the four published ACOX1 IHC protocols below provide starting points for paraffin sections (page protocol; PMC11757735; PMC9453879; PMC10154930; PMC10601207).
| Sample | FFPE human colon carcinoma tissue (datasheet M03054) |
| Fixation | Image formalin-fixed; duration unreported (datasheet M03054); 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 | Mouse monoclonal (clone 153CT43.1.1) anti-ACOX1, 1:10-1:50 (datasheet M03054) |
| 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 | ACOX1-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page retrieval); use EDTA when reproducing PMC10601207.
Section 2
What Is the Expected ACOX1 Staining Pattern?
ACOX1 is a peroxisomal protein with no annotated transmembrane segment (UniProt Q15067). In paraffin sections, expect granular cytoplasmic staining, especially in hepatocytes and duodenal or small-intestinal glandular cells (HPA: tissue IHC). HPA rates the tissue staining “Approved” but reports low consistency with RNA expression data (HPA: tissue IHC reliability); interpret intensity with that limitation in mind.
| Fine cytoplasmic granules in hepatocytes or intestinal glandular cells. | This matches the expected peroxisomal distribution and the reported High staining in these cells (UniProt Q15067; HPA: tissue IHC). Compare the granules with local background before scoring. |
| Predominant cell-surface, diffuse nuclear, or other non-granular staining. | These patterns do not match the general granular cytoplasmic tissue pattern (HPA: tissue IHC). ACOX1 has no annotated transmembrane segment (UniProt Q15067). Check whether the signal follows tissue edges or detection background before calling it ACOX1. |
| Strong staining in a cell population expected to stain only weakly. | For example, strong skeletal-muscle myocyte staining conflicts with HPA’s Low observation (HPA: skeletal muscle). Consider antibody cross-reactivity or endogenous detection activity; a low-staining tissue is not a validated negative control. |
| Uniform haze over cells, stroma, and empty areas. | A broad haze obscures the granular cytoplasmic pattern reported for ACOX1 (HPA: tissue IHC). In general IHC practice, incomplete blocking, concentrated primary antibody, or detection background can produce such staining; assess it with a primary-omission control. |
| No convincing signal in hepatocytes on an otherwise intact section. | Hepatocytes are reported as High in HPA tissue IHC (HPA: liver). First check tissue identity and assay controls, then review the IHC-validated antibody’s retrieval, dilution, and detection settings; this result alone does not establish absent ACOX1. |
💡Expected ACOX1 appearanceCall a positive result when High, granular cytoplasmic staining is seen in hepatocytes or duodenal or small-intestinal glandular cells (HPA: tissue IHC; UniProt Q15067 peroxisome); widespread haze or dominant surface staining is suspect because it does not match that reported pattern (HPA: tissue IHC; UniProt Q15067 topology).
| Tissue selection and scoring | HPA reports High staining in liver hepatocytes and duodenal and small-intestinal glandular cells, with Low staining in several other cell types (HPA: tissue IHC). Score the named cells and their granular pattern, rather than assigning a single intensity to the whole section. |
| Isoforms across tissues | UniProt lists 3 isoforms and reports different relative levels of isoforms 1 and 2 across tissues (UniProt Q15067). Tissue-to-tissue intensity differences therefore need context; the supplied sources do not establish which isoforms any IHC antibody detects. |
| Processing and epitope coverage | UniProt lists a 1–660 chain and processed 1–468 and 469–660 chains (UniProt Q15067). An antibody’s epitope position could affect recognition of those chains; no epitope location is supplied here, so chain-specific staining cannot be inferred. |
| Strength of tissue evidence | HPA labels its ACOX1 tissue IHC profile Approved while reporting low consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). HPA also lists four antibodies with Approved IHC status (HPA: antibody validation); those labels do not resolve an unexpected result in an individual section. |
| IF/ICC Q&A: What localisation should appear? | The main IF/ICC location is peroxisomes, with an additional nucleolar location (HPA: subcellular ICC-IF). Treat that nucleolar observation as IF/ICC evidence; the supplied tissue IHC profile describes a general granular cytoplasmic pattern (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Granules are absent from liver hepatocytes. | The result conflicts with HPA’s High hepatocyte observation (HPA: liver); tissue or assay failure is possible. | Confirm hepatocyte-rich tissue and working detection controls. Review the IHC-validated antibody’s stated retrieval and dilution settings before interpreting a negative result (general IHC practice). |
| Chromogen covers the section evenly. | Uniform colour is unlike HPA’s granular cytoplasmic profile (HPA: tissue IHC); non-specific detection background is possible. | Inspect a primary-omission control and review blocking, washes, and primary concentration (general IHC practice). If using an enzyme reporter, check for matching endogenous enzyme activity. |
| Signal follows edges or appears mainly on cell surfaces. | That distribution conflicts with the reported granular cytoplasm and lacks support from ACOX1 topology (HPA: tissue IHC; UniProt Q15067). | Compare with adjacent intact tissue and a primary-omission control. Reassess whether the signal follows section damage or detection background (general IHC practice). |
| Unexpected cells stain more strongly than hepatocytes. | Cell identity or assay background may be misleading; HPA reports High hepatocytes but also notes low staining-to-RNA consistency (HPA: tissue IHC). | Score identified cell types separately and compare with a known-positive liver section and a primary-omission control (HPA: liver; general IHC practice). Do not use intensity alone to assign specificity. |
| A low-staining tissue shows no visible signal. | HPA lists Low staining for cell types including skeletal-muscle myocytes and salivary-gland cells (HPA: tissue IHC). | Check a high-staining liver or intestinal control before judging assay performance (HPA: tissue IHC). Do not treat a low-staining cell type as a proven ACOX1-negative control. |
| IF/ICC shows nucleolar signal alongside puncta. | HPA reports peroxisomes as the main IF/ICC location and nucleoli as an additional location (HPA: subcellular ICC-IF). | Interpret the IF/ICC observation in its own assay context. For paraffin IHC, assess the reported granular cytoplasmic tissue pattern and appropriate controls (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ACOX1 staining
M03054 · IHCACOX1 Monoclonal Antibody immunohistochemistry analysis in formalin fixed and paraffin embedded human colon carcinoma followed by peroxidase
M03054 · IFConfocal immunofluorescent analysis of ACOX1 Antibody with Hela cell followed by Alexa Fluor® 488-conjugated goat anti-mouse lgG (green). DA
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000161533-ACOX1/tissue/cerebral+cortex#imid_5758900
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000161533-ACOX1/tissue/colon#imid_5758826
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000161533-ACOX1/tissue/liver#imid_5758811
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000161533-ACOX1/tissue/kidney#imid_5758820
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021195; green = ACOX1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021195; green = ACOX1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACOX1 IHC & IF
🧪Run liver first: hepatocytes should stain strongly (HPA: High in hepatocytes). HPA detects ACOX1 in all 45 scored tissues, so there is no HPA-supported negative tissue; use no-primary and isotype controls, and treat cells without specific staining on the liver slide as background references rather than proven ACOX1-negative cells (HPA: no negative rows; detected in all 45 tissues).
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: None in HPA: ACOX1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ACOX1 in A-431, U-251MG, U2OS, with annotated localisation: Peroxisomes (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a control with a host- and isotype-matched monoclonal antibody; an ACOX1 knockout sample with confirmed target loss provides a biological negative (selected caption: monoclonal antibody; standard IHC practice). For liver chromogenic IHC, quench endogenous peroxidase and assess pigment background before interpreting DAB staining (HPA: liver hepatocytes High; standard IHC practice).
⚠️Feasibility: The selected catalog antibody caption reports staining in formalin-fixed, paraffin-embedded colon carcinoma, but supplies no target-specific fixation window or retrieval condition (selected M03054 tissue-IHC caption). Whether ACOX1 staining depends on antigen retrieval, or whether frozen sections are easier, is unreported in the supplied evidence. IF/ICC can help assess the expected punctate peroxisomal pattern, although its ease relative to IHC is unestablished; liver pigment and endogenous peroxidase can complicate chromogenic interpretation (UniProt Q15067: peroxisome; HPA: peroxisomes approved; standard IHC practice).
HPA tissue IHC evidence for ACOX1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ACOX1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ACOX1 IHC Tips
Troubleshoot ACOX1 chromogenic IHC in paraffin sections by checking retrieval, staining compartment, controls, and scoring before interpreting a granular signal.
How should I retrieve ACOX1 in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). After cooling and washing, compare a known positive tissue section with a no-primary control in the same run; liver hepatocytes or duodenal glandular cells provide documented strong staining references (HPA: High in hepatocytes and duodenal glandular cells). If signal remains weak, test a more stringent retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Judge improvement by sharper granular cytoplasmic staining, since broad diffuse color alone does not match the reported tissue pattern (HPA: general cytoplasmic granular expression).
Does formalin fixation explain weak or uneven ACOX1 staining?
The catalog antibody M03054 has been shown on formalin-fixed, paraffin-embedded human colon carcinoma with peroxidase detection and DAB, but the caption provides no comparison of fixation conditions (M03054 tissue-IHC caption). ACOX1-specific sensitivity to fixation is therefore unknown; do not infer it from its peroxisomal location, processing, or tissue distribution (UniProt Q15067: peroxisome and processed chains; HPA: tissue profile). Review fixation records and compare sections processed together, because inconsistent fixation can change antigen access in routine IHC (standard IHC practice). Keep retrieval, staining time, and detection conditions matched, then use an internal tissue reference and no-primary control to separate processing variation from nonspecific color (standard IHC practice).
Where should convincing ACOX1 staining appear in an IHC section?
Expect a fine granular cytoplasmic signal consistent with a peroxisomal protein; ACOX1 has no annotated transmembrane segment (HPA: general cytoplasmic granular expression; UniProt Q15067: peroxisome and topology). Assess granules within identifiable cells rather than treating uniform cytoplasmic haze or cell borders as equivalent evidence (HPA: tissue IHC profile; standard IHC practice). Liver hepatocytes and duodenal glandular cells are useful tissue references because their recorded staining is High (HPA: High in hepatocytes and duodenal glandular cells). HPA also reports an approved nucleolar location in cell imaging, so apparent nuclear IHC signal needs separate control and compartment assessment before assignment to ACOX1 (HPA: approved nucleolar location).
Could isoforms or ACOX1 processing change what the antibody detects?
ACOX1 has 3 annotated isoforms, and its recorded processing includes a full-length A chain plus B and C chains, so epitope placement matters when interpreting section staining (UniProt Q15067: isoforms and processing). Without an epitope map for the catalog antibody, do not assume that every isoform or processed chain is recognized equally (UniProt Q15067: isoforms and processing; antibody epitope unspecified in supplied evidence). Isoform 1 exceeds isoform 2 in liver and kidney, whereas isoform 2 is higher in brain, lung, muscle, white adipose tissue, and testis (UniProt Q15067: tissue specificity). Compare tissues only after matching processing and scoring, and interpret discordance as an epitope question rather than proof of altered protein abundance (standard IHC practice).
How can IF help verify a suspicious ACOX1 IHC pattern?
On a separate IF/ICC preparation, multiplex ACOX1 with a marker identifying the expected cell population, such as a hepatocyte marker when assessing liver, and compare signals cell by cell (HPA: High in hepatocytes; standard IF practice). Choose fluorophores after checking tissue autofluorescence in unstained controls, favoring channels with cleaner background for the weaker signal (standard IF practice). Permeabilise sufficiently for antibody access to the cytosolic face of peroxisomes, while checking the antibody epitope because its location is unspecified here and ACOX1 has no annotated transmembrane segment (UniProt Q15067: peroxisome and topology; standard IF practice). Assess puncta against a peroxisomal reference marker, while recognizing that IF/ICC optimization does not establish performance in chromogenic paraffin IHC (HPA: approved peroxisomal location; standard IF practice).
What should I check when ACOX1 DAB staining looks diffuse or widespread?
First compare the stained section with a no-primary control, because the expected tissue profile is granular cytoplasmic staining rather than uniform brown haze (HPA: general cytoplasmic granular expression; standard IHC practice). In a peroxidase and DAB workflow, confirm endogenous peroxidase blocking and inspect pigment or precipitate before assigning brown material to ACOX1 (M03054 tissue-IHC caption: peroxidase and DAB; standard IHC practice). Check whether excess primary antibody, incomplete washing, or uneven drying tracks the background across the section (standard IHC practice). Use strongly stained liver hepatocytes or duodenal glandular cells as positive references, while treating low-staining salivary glandular cells as a useful comparison rather than an absolute negative (HPA: High in liver and duodenum; Low in salivary gland).
How should I score heterogeneous ACOX1 staining across a specimen? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of cells with granular cytoplasmic staining and an intensity category for each field (HPA: general cytoplasmic granular expression; standard IHC scoring practice). An H-score can combine percentages across intensity categories on a 0–300 scale; report the scoring thresholds alongside the result (standard IHC scoring practice). For spatial questions, measure positive-cell density per mm² of viable tissue and normalize comparisons to the same cell type, evaluable area, section handling, and detection run (standard IHC scoring practice). Keep liver hepatocytes or duodenal glandular cells as run references, but do not use their High designation as a universal numeric cutoff (HPA: High in hepatocytes and duodenal glandular cells).
When is an ACOX1-positive IHC result biologically credible?
A credible result shows reproducible granular cytoplasmic staining in intact cells, consistent with peroxisomal ACOX1 and the reported tissue pattern (UniProt Q15067: peroxisome; HPA: general cytoplasmic granular expression). Compare the observed cell type with documented High staining in liver hepatocytes or duodenal glandular cells, while remembering that the tissue-IHC record reports low consistency between staining and RNA expression (HPA: High cell types; HPA: reliability description). Treat edge-only staining, necrotic areas, diffuse nuclear color, and signal retained in a no-primary control as artefact candidates; assess endogenous peroxidase when DAB is used (standard IHC practice; M03054 tissue-IHC caption: DAB). The catalog image demonstrates staining in formalin-fixed, paraffin-embedded human colon carcinoma, but its caption states that clinical relevance has not been evaluated (M03054 tissue-IHC caption).
Boster reagents
Best ACOX1 / Peroxisomal acyl-coenzyme A oxidase 1 IHC Antibodies
Anti-ACOX1 catalog data include human colon carcinoma IHC and HeLa cell IF images (M03054 image captions), plus human, mouse, and rat reactivity for A03054-1 (catalog).
Real IHC data
Anti-ACOX1 Antibody
Cat # M03054
M03054 is the figure-backed card: its captions show DAB IHC in formalin-fixed, paraffin-embedded human colon carcinoma and confocal IF in HeLa cells (M03054 image captions). A03054-1 lists human IHC and ICC/IF applications but has no IHC or IF image in the payload, so it has no figure-backed card (A03054-1 catalog).
Which to pick: For human tissue IHC, choose M03054: it is a mouse monoclonal listed for IHC-P, with a formalin-fixed, paraffin-embedded human colon carcinoma example (M03054 catalog; M03054 IHC image caption). For human cell IF, M03054 has a HeLa image; A03054-1 is a rabbit polyclonal listed for human ICC/IF without an accompanying image (M03054 IF image caption; A03054-1 catalog). For cross-species planning, A03054-1 lists human, mouse, and rat reactivity, but its paraffin IHC listing specifies human, and no IHC image or fixative is reported for it (A03054-1 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15067 (ACOX1_HUMAN, Peroxisomal acyl-coenzyme A oxidase 1).
- Human Protein Atlas. ACOX1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ACOX1 subcellular location (ICC-IF): Mainly localized to the peroxisomes. In addition localized to the nucleoli..
- Human Protein Atlas. ACOX1 antibody validation summary (4 antibodies).
- ACOX1 activates autophagy via the ROS/mTOR pathway to suppress proliferation and migration of colorectal cancer. Scientific reports 2025 — PMC11757735.
- Bailing capsule (Cordyceps sinensis) ameliorates renal triglyceride accumulation through the PPARα pathway in diabetic rats. Frontiers in pharmacology 2022 — PMC9453879.
- ACOX-driven peroxisomal heterogeneity and functional compartmentalization in brown adipocytes of hypothyroid rats. Royal Society open science 2023 — PMC10154930.
- Silencing of CD147 inhibits cell proliferation, migration, invasion, lipid metabolism dysregulation and promotes apoptosis in lung adenocarcinoma via blocking the Rap1 signaling pathway. Respiratory research 2023 — PMC10601207.
- PubMed PMID:8159712 — UniProt-cited evidence.
- PubMed PMID:7876265 — UniProt-cited evidence.
- PubMed PMID:8040306 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACOX1 (Peroxisomal acyl-coenzyme A oxidase 1). Expected localisation Granular cytoplasm (HPA tissue IHC); peroxisome (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M03054 at 1:10-1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: FFPE human colon carcinoma tissue (datasheet M03054); 2. Fixation: Image formalin-fixed; duration unreported (datasheet M03054); 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: Mouse monoclonal (clone 153CT43.1.1) anti-ACOX1, 1:10-1:50 (datasheet M03054); 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: Staining has low concordance with tissue RNA (HPA tissue IHC); Isoform / epitope: 3 isoforms; B/C processing may alter epitope detection (UniProt); Regulation: Liver RNA is tissue enhanced (HPA tissue RNA). Evidence: UniProt Q15067, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the peroxisomes. In addition localized to the…, Datasheet M03054, PMC11757735, PMC9453879. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACOX1/immunohistochemistry-ihc-guide.