ACOX1 / Peroxisomal acyl-coenzyme A oxidase 1 · IHC design guide

Design Immunohistochemistry for ACOX1

Plan chromogenic ACOX1 IHC-P with catalog antibody M03054 at 1:10–1:50 (datasheet: M03054). Use high-staining hepatocytes as a positive tissue reference and assess granular cytoplasmic staining (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for ACOX1 (IHC for ACOX1): expected localisation Granular cytoplasm (observed); peroxisome (molecular) (HPA tissue IHC; UniProt), antibody M03054, validated IHC image, and IHC protocol steps — Printable **ACOX1** IHC protocol sheet — expected localisation Granular cytoplasm (observed); peroxisome (molecular) (HPA tissue IHC; UniProt), antibody M03054, controls and protocol steps. Open the full ACOX1 IHC guide →

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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 (observed); peroxisome (molecular) (HPA tissue IHC; UniProt)
	Staining pattern	Granular cytoplasmic staining across tissues (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 samples. (standard IHC practice; not target-specific)
	Caveat	Antibody staining has low consistency with RNA expression (HPA tissue IHC)
	Regulation	Isoform abundance varies by tissue (UniProt)
	Isoform / epitope	3 isoforms; B/C chains split at 468/469—check epitope (UniProt)

Section 1

Recommended ACOX1 IHC & IF Protocols

The catalog antibody protocol is accompanied by published ACOX1 IHC methods for colorectal cancer sections, rat kidney, rat brown adipose tissue, and tumor xenografts (PMC11757735; PMC9453879; PMC10154930; PMC10601207).

Recommended immunohistochemistry (IHC-P) protocol parameters
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, the page’s specified condition and a reported approach in two protocols; try EDTA if staining needs optimization (PMC11757735; PMC10154930; PMC10601207).

Section 2

What Is the Expected ACOX1 Staining Pattern?

ACOX1 is a peroxisomal protein with no transmembrane segment (UniProt Q15067 topology and subcellular location). In paraffin-section IHC, expect granular cytoplasmic staining, especially in liver hepatocytes and duodenal and small-intestinal glandular cells (HPA: tissue IHC profile; High in these cells). HPA rates its tissue staining Approved but reports low consistency between antibody staining and RNA expression, so interpret intensity with that limitation (HPA: reliability description).

What am I looking at on my slide?
Granular cytoplasmic staining in hepatocytes or intestinal glandular cells.	This fits the reported tissue pattern and High staining in those cells (HPA: tissue IHC). Granules are consistent with peroxisomal localization (UniProt Q15067; HPA: subcellular). Brightfield IHC does not resolve every granule as an individual peroxisome (general IHC practice).
Predominantly membranous or nuclear staining, with little granular cytoplasmic signal.	Question target assignment: ACOX1 has no transmembrane segment and is peroxisomal (UniProt Q15067 topology and location). Nucleolar localization is additionally reported by ICC-IF, so nucleolar signal alone cannot be declared an artefact, but it does not replace the expected tissue IHC pattern (HPA: subcellular; tissue IHC).
Strong staining in unexpected cells while nearby expected cells show little signal.	Consider cross-reactivity or endogenous detection activity, then compare morphology and controls (general IHC practice). HPA reports widespread cytoplasmic expression and lists no negative tissues; an unlisted cell type is not automatically ACOX1-negative (HPA: tissue IHC).
Uniform haze across tissue, or color in tissue-free areas.	A field-wide signal lacks the reported granular cytoplasmic pattern (HPA: tissue IHC). Background from detection reagents, nonspecific binding, or incomplete washing can obscure cell-level interpretation; assess reagent controls and washing before scoring (general IHC practice).
No convincing signal in liver hepatocytes or intestinal glandular cells.	These are reported High-staining cells, so first check section quality, retrieval and detection controls (HPA: tissue IHC; general IHC practice). A failed run is possible, but the HPA reliability caveat means one unstained specimen does not by itself settle antibody specificity or biological absence (HPA: reliability description).

💡Expected ACOX1 appearanceCall a positive result when hepatocytes or duodenal or small-intestinal glandular cells show High granular cytoplasmic staining (HPA: tissue IHC); predominant membrane staining or uniform field-wide color is suspect (UniProt Q15067 topology; general IHC practice).

How each factor affects the staining
Compartment and optical resolution	ACOX1 is peroxisomal (UniProt Q15067; HPA: subcellular). The tissue IHC readout is granular cytoplasm (HPA: tissue IHC); do not require every granule to be separately resolved in chromogenic sections (general IHC practice).
Tissue and cell selection	HPA reports High staining in liver hepatocytes and duodenal and small-intestinal glandular cells, and Low staining in several listed cell types (HPA: tissue IHC). Use the High group as practical positive references; Low is not a validated absence control (HPA: tissue IHC).
Isoforms and processing	UniProt lists 3 isoforms and full-length, B-chain and C-chain annotations; isoform balance varies by tissue (UniProt Q15067). Without an antibody epitope in the payload, no isoform or chain-specific IHC interpretation is justified (UniProt Q15067; general antibody interpretation).
Antibody evidence	HPA lists four antibodies with Approved IHC status, while its tissue profile reports low staining-to-RNA consistency (HPA: antibody statuses; reliability description). Approved supports using the observations as a reference, but does not establish that every granular signal in a new run is specific (general IHC practice).
IF/ICC Q&A: should the same pattern appear?	In ICC-IF, HPA reports peroxisomes as the main location and nucleoli as an additional location, with images from A-431, U-251MG and U2OS (HPA: subcellular). This supports a localization comparison, not an IF protocol or an expectation that nucleolar tissue IHC must be visible (HPA: subcellular; general microscopy practice).
Detection chemistry	Endogenous enzyme activity or nonspecific secondary-reagent binding can produce color unrelated to ACOX1 (general IHC practice). A reagent control helps identify that contribution; the supplied HPA and UniProt records do not quantify its size in any listed tissue.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Positive-reference cells show no signal.	The run may have insufficient retrieval or a detection failure (general IHC practice); target-specific fixation sensitivity is unreported in the supplied records.	Check a contemporaneous positive section and detection controls, then adjust retrieval within the chosen IHC workflow (general IHC practice). Use hepatocytes or intestinal glandular cells as reported High references (HPA: tissue IHC).
Granules are faint but tissue-wide haze is strong.	Background may mask the granular cytoplasmic pattern reported by HPA (HPA: tissue IHC; general IHC practice).	Review reagent-only controls, blocking, primary-antibody concentration and wash steps; compare cells against adjacent background before scoring (general IHC practice). No ACOX1-specific dilution is supplied.
Signal outlines cell membranes.	A predominantly membrane pattern conflicts with peroxisomal localization and the absence of a transmembrane segment (UniProt Q15067 topology; HPA: subcellular).	Check morphology and control sections, then repeat with the IHC-validated antibody and its documented conditions if needed (general IHC practice). Do not score membrane-only color as the expected ACOX1 pattern.
A surprising cell population stains strongly.	Cross-reactivity or endogenous detection activity is possible (general IHC practice), but ACOX1 tissue expression is broad and HPA supplies no negative-tissue list (HPA: tissue IHC).	Compare the compartment and staining pattern with nearby expected cells; inspect reagent controls before assigning specificity or declaring the population biologically negative (HPA: tissue IHC; general IHC practice).
Nucleolar signal appears in an IF comparison.	HPA reports additional nucleolar localization in ICC-IF alongside the main peroxisomal location (HPA: subcellular).	Assess whether peroxisomal signal is also present and keep the ICC-IF observation separate from tissue IHC scoring (HPA: subcellular; tissue IHC). Follow the separate IF/ICC guide for experimental conditions.
Two specimens have different staining intensities.	HPA reports tissue-dependent staining levels, and UniProt reports tissue-dependent isoform balance; HPA also notes low consistency with RNA expression (HPA: tissue IHC; UniProt Q15067 tissue specificity).	Compare matched cell types and run controls under the same IHC conditions before calling a biological change; do not infer a particular isoform or chain from intensity alone (general IHC practice; UniProt Q15067).

Source: UniProt Q15067 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ACOX1 staining

ACOX1 Monoclonal Antibody immunohistochemistry analysis in formalin fixed and paraffin embedded human colon carcinoma followed by peroxidase conjgation of the secondary antibody and DAB staining. This data demonstrates the use of the ACOX1 Monoclonal Antibody for immunohistochemistry. Clinical relevance has not been evaluated.

M03054 · IHCACOX1 Monoclonal Antibody immunohistochemistry analysis in formalin fixed and paraffin embedded human colon carcinoma followed by peroxidase

Confocal immunofluorescent analysis of ACOX1 Antibody with Hela cell followed by Alexa Fluor® 488-conjugated goat anti-mouse lgG (green). DAPI was used to stain the cell nuclear (blue).

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.

← 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 (HPA: High in hepatocytes). HPA detects ACOX1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect cells without detectable signal on the positive slide to show counterstain alone rather than specific punctate staining (HPA: no negative tissue listed; UniProt Q15067: peroxisome).

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 slide; an isotype control matched to the catalog antibody’s host species and immunoglobulin class; and ACOX1 knockout material as a biological negative (selected-SKU caption: monoclonal antibody; standard IHC practice). Quench endogenous peroxidase in liver before peroxidase/DAB detection (selected-SKU caption: peroxidase/DAB; standard IHC practice).

⚠️Feasibility: The selected-SKU caption reports formalin-fixed, paraffin-embedded human colon carcinoma stained with peroxidase/DAB, but reports no target-specific fixation window or fixation effect (selected-SKU caption: M03054). Antigen-retrieval dependence is unreported; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU caption; HPA: ICC-IF images available). In liver, endogenous peroxidase may add DAB background, so assess staining against the no-primary control (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 staining by checking retrieval, tissue context and peroxisomal localisation before comparing chromogenic IHC results across sections.

How should I retrieve ACOX1 in paraffin sections when staining is weak?

Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval rule). Apply the same cooling interval and staining run to test and control sections so retrieval differences do not masquerade as expression changes (standard IHC practice). The selected antibody has been used on formalin fixed, paraffin embedded human colon carcinoma with peroxidase and DAB detection, but its caption does not specify a retrieval method (M03054 tissue-IHC caption). If staining remains weak, compare a more alkaline retrieval buffer on adjacent sections as a controlled fallback, recording any increase in diffuse background (standard IHC practice).

Could fixation explain inconsistent ACOX1 staining between paraffin blocks?

The selected antibody's tissue image used formalin fixed, paraffin embedded human colon carcinoma (M03054 tissue-IHC caption). No supplied evidence establishes ACOX1-specific fixation sensitivity, so treat fixation time as a variable to test rather than an established cause of signal loss (supplied fixation evidence). Compare sections with documented fixation histories using the same pH 6.0, 20 min retrieval and one detection run (page retrieval rule; standard IHC practice). Include a known staining control on each run, and assess preserved morphology alongside signal before changing antibody concentration (standard IHC practice). Do not infer fixation behaviour from the protein's modifications or its observed tissue pattern (UniProt ACOX1 record; HPA tissue IHC).

Where should convincing ACOX1 signal appear in chromogenic IHC?

Expect cytoplasmic granularity rather than a smooth nuclear or membrane outline: tissue IHC reports a granular cytoplasmic pattern, and ACOX1 is peroxisomal (HPA tissue IHC; UniProt ACOX1 subcellular location). Compare hepatocytes or intestinal glandular cells with nearby lower staining areas within a section, while preserving their identities during scoring (HPA: High in hepatocytes and intestinal glandular cells; standard IHC practice). ACOX1 has no annotated transmembrane segment, so an isolated crisp plasma membrane rim needs further validation (UniProt ACOX1 topology). Nuclear signal requires caution: approved ICC/IF localisation includes nucleoli, but that observation alone does not validate nuclear DAB staining in paraffin tissue (HPA subcellular; standard IHC practice).

Can this stain distinguish ACOX1 isoforms or processed chains?

Do not assign an isoform from chromogenic intensity alone: the record lists 3 isoforms, while the supplied tissue-IHC caption gives no epitope or isoform selectivity (UniProt ACOX1 isoforms; M03054 tissue-IHC caption). Isoform abundance also varies by tissue, which can complicate comparisons even when staining is technically consistent (UniProt ACOX1 tissue specificity). The record describes a 1–660 chain and shorter 1–468 and 469–660 chains; epitope position therefore matters when interpreting a negative stain (UniProt ACOX1 processing). Obtain an epitope map or independent isoform evidence before claiming isoform-specific or chain-specific localisation, and keep scoring labels at the ACOX1 level meanwhile (standard IHC practice).

How can IF help assess an ambiguous ACOX1 IHC pattern?

On a separate IF preparation, pair ACOX1 with a marker for the expected cell type, such as a hepatocyte marker when examining liver, and inspect signal within identified cells (HPA: High in hepatocytes; standard IF practice). Choose spectrally separated fluorophores and consider a far-red ACOX1 channel if tissue autofluorescence obscures shorter wavelengths (standard IF practice). ACOX1 is peroxisomal with no annotated transmembrane segment, so permeabilise for an intracellular epitope and titrate detergent to preserve punctate structures (UniProt ACOX1 location and topology; standard IF practice). Compare puncta with a peroxisomal marker; HPA also reports nucleolar localisation in ICC/IF, which warrants separate assessment before transferring that interpretation to chromogenic IHC (HPA subcellular).

What should I check when ACOX1 DAB staining looks diffuse?

First compare the diffuse signal with the expected granular cytoplasmic pattern in tissue sections (HPA tissue IHC). Run a no-primary control, apply a peroxidase block before DAB development, and check whether pigment or endogenous enzyme activity persists without primary antibody (standard chromogenic IHC practice). The selected tissue image used a peroxidase-conjugated secondary and DAB, but it supplies no ACOX1-specific background or blocking result (M03054 tissue-IHC caption). If controls are clean, titrate primary antibody and DAB development on adjacent sections while holding retrieval at pH 6.0 for 20 min (page retrieval rule; standard IHC practice). Evaluate cellular granularity and overall background together before selecting conditions (standard IHC practice).

How should I score ACOX1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring; ACOX1 tissue staining is generally granular and cytoplasmic (HPA tissue IHC). Within that region, record the percentage of positive cells and intensity to calculate an H-score, or measure positive granular area per mm² of viable tissue (standard IHC quantification practice). Normalise to the same eligible cell population or viable tissue area, and apply identical thresholds, illumination and DAB development criteria across the comparison (standard IHC quantification practice). Report retrieval and staining batches, since a shift in processing can change measured intensity (standard IHC practice). Interpret small differences cautiously because HPA labels its tissue IHC approved yet reports low consistency with RNA expression (HPA tissue IHC).

How do I distinguish true ACOX1 staining from artefact?

Look for granular cytoplasmic staining in intact cells and compare it with expected peroxisomal localisation (HPA tissue IHC; UniProt ACOX1 subcellular location). Signal in hepatocytes or intestinal glandular cells has tissue-context support, whereas a smooth membrane rim lacks support from the recorded topology (HPA: High in hepatocytes and intestinal glandular cells; UniProt ACOX1 topology). Inspect edges and necrotic regions separately, and reject staining that tracks tissue damage or appears in the no-primary control (standard IHC practice). Check a peroxidase-blocked control when brown deposit could reflect endogenous enzyme activity (standard chromogenic IHC practice). Treat nucleolar staining as unresolved in paraffin IHC even though HPA reports it in ICC/IF (HPA subcellular).

Boster reagents

Best ACOX1 / Peroxisomal acyl-coenzyme A oxidase 1 IHC Antibodies

M03054 has IHC data from formalin-fixed, paraffin-embedded human colon carcinoma and IF data from HeLa cells (catalog IHC and IF captions).

Real IHC data ACOX1 Monoclonal Antibody immunohistochemistry analysis in formalin fixed and paraffin embedded human colon carcinoma followed by peroxidase conjgation of the secondary antibody and DAB staining. This data demonstrates the use of the ACOX1 Monoclonal Antibody for immunohistochemistry. Clinical relevance has not been evaluated.

Anti-ACOX1 Antibody

Cat # M03054

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M03054 is the card that will render; its IHC image shows peroxidase/DAB staining of formalin-fixed, paraffin-embedded human colon carcinoma (catalog IHC caption). Its IF image shows staining of HeLa cells, and its listed reactivity is human (catalog IF caption; catalog reactivity).

Which to pick: Choose mouse monoclonal M03054 for human tissue IHC-P because its own image documents formalin-fixed, paraffin-embedded colon carcinoma stained with peroxidase/DAB (catalog host/clone; catalog IHC caption). M03054 also has an IF image in HeLa cells; rabbit polyclonal A03054-1 lists both IF and ICC (catalog IF caption; catalog host/applications). For mouse or rat samples, consider A03054-1 because those species are listed as reactive, but its stated paraffin-section IHC condition is for human samples (catalog reactivity; datasheet: IHC dilution).

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.