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ACOT13 / Acyl-coenzyme A thioesterase 13 · IHC design guide
Design Immunohistochemistry for ACOT13
Plan chromogenic ACOT13 IHC in paraffin sections using liver hepatocytes or kidney tubule cells as positive tissue references (HPA tissue IHC). The catalog antibody starts at 2.5 μg/mL (datasheet); assess granular cytoplasmic staining against a negative control (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACOT13 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 in tissue sections (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm in hepatocytes and kidney tubule cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Gallbladder+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07288-1) | |
| Caveat | Antibody–RNA consistency is medium (HPA tissue IHC) | |
| Regulation | No regulatory stimulus specified (UniProt record) | |
| Isoform / epitope | 2 isoforms; N-terminal processing may affect N-terminal epitopes (UniProt) |
Section 1
Recommended ACOT13 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published rat caudal intervertebral disc protocol (PMC12983563 methods).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07288-1); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-ACOT13, 2.5 μg/mL (datasheet A07288-1) |
| 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 | ACOT13-positive staining in glandular cells of gallbladder (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min (page retrieval rule); the rat disc study used citrate retrieval (PMC12983563 methods).
Section 2
What Is the Expected ACOT13 Staining Pattern?
ACOT13 should show chiefly granular cytoplasmic staining in tissue sections (HPA tissue IHC: general cytoplasmic expression with a granular pattern). Strong examples include hepatocytes, kidney tubule cells and gallbladder glandular cells (HPA tissue IHC: High). UniProt also lists cytosol, mitochondria, nucleus and mitotic spindle, and reports no transmembrane segment (UniProt Q9NPJ3). Interpret compartment differences cautiously: HPA rates tissue staining Enhanced, with medium consistency against RNA data (HPA tissue IHC: reliability).
| Granular cytoplasmic staining in hepatocytes or kidney tubule cells, with clear cell outlines. | This matches the reported tissue pattern and High staining in those cells (HPA tissue IHC). Compare the named cell population and its cytoplasmic distribution; intensity elsewhere need not match it (HPA tissue IHC: tissue-specific levels). |
| Staining appears exclusively nuclear, or as a continuous rim around most cells. | Neither is the reported general tissue IHC pattern (HPA tissue IHC: granular cytoplasm). UniProt does list nuclear ACOT13 during interphase, so nuclear signal alone cannot establish an artefact (UniProt Q9NPJ3). Check morphology and controls before assigning localisation (general IHC practice). |
| Strong staining dominates adipocytes, smooth muscle cells or oral squamous epithelial cells. | Those populations are listed as Not detected in the HPA tissue examples (HPA tissue IHC). Unexpected signal raises cross-reactivity or detection-background questions; it does not establish either cause by itself. Review cell identity and control sections (general IHC practice). |
| Color spreads across extracellular areas or the whole section without discernible cellular boundaries. | That appearance cannot be scored as the reported granular cytoplasmic pattern (HPA tissue IHC). Excess detection background or endogenous activity can produce broad color in chromogenic IHC; inspect a negative reagent control and section pigment (general IHC practice). |
| Hepatocytes remain unstained in an otherwise readable section. | Hepatocytes are a High HPA tissue example (HPA tissue IHC), so absent signal calls for checking that the positive control and detection run worked. It does not, on its own, show that the specimen lacks ACOT13 (general IHC practice). |
💡Expected ACOT13 appearanceCall a section positive when the named cells show discernible granular cytoplasmic color, especially High-level hepatocytes or kidney tubule cells (HPA tissue IHC); broad extracellular color or strong signal in HPA Not detected cells warrants a background check, rather than a positive call (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Hepatocytes, kidney tubule cells, gallbladder glandular cells, ovary follicle cells, parathyroid glandular cells and late spermatids are High examples (HPA tissue IHC). Score the specified cells within each tissue; HPA levels are observations, not a guarantee for every section. |
| Compartment evidence | Tissue IHC is generally granular and cytoplasmic (HPA tissue IHC). UniProt additionally lists nucleus and mitotic spindle, including interphase nuclear localisation and mitotic tubulin colocalisation (UniProt Q9NPJ3). These annotations do not make nuclear-only tissue staining the expected IHC result. |
| Antibody validation | HPA019881 and HPA057134 each have Enhanced IHC status (HPA antibodies). HPA describes Enhanced as reproduction by independent antibodies or orthogonal data; the tissue summary still reports medium consistency with RNA expression (HPA tissue IHC). Interpret a new reagent against its own controls. |
| Isoforms and processing | UniProt lists 2 isoforms and chains beginning at residue 1 or 2, including an N-terminally processed form (UniProt Q9NPJ3). No antibody epitope is supplied here, so these facts cannot predict whether the catalog antibody stains the forms differently. |
| IF/ICC Q&A | What if IF shows junctional signal? HPA ICC-IF reports mainly cell junction localisation and marks it Approved, with images in A-431 and U-251MG (HPA subcellular). That separate observation does not replace the granular cytoplasmic expectation for tissue IHC (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No color in the ACOT13 positive control. | A failed detection run or unsuitable staining conditions are possible (general IHC practice); HPA reports High hepatocyte and kidney tubule staining (HPA tissue IHC). | Confirm tissue identity, section integrity, reagent sequence and detection-control performance; review the catalog antibody's IHC-P instructions before changing conditions (general IHC practice). |
| Positive cells show faint, hard-to-score color. | HPA levels vary by cell population; cerebral cortical neurons and lung macrophages are Low examples (HPA tissue IHC). | Score a High example such as hepatocytes beside the test section, then assess cell-specific color and background on the same run (HPA tissue IHC; general IHC practice). |
| Color covers tissue and empty areas alike. | Detection background or endogenous chromogenic activity may obscure cellular staining (general IHC practice). | Inspect a control without primary antibody and any native pigment; check blocking, washes and detection timing using the established IHC workflow (general IHC practice). |
| Unexpected strong color appears in HPA Not detected cells. | Cross-reactivity, endogenous activity or mistaken cell identification are possibilities; HPA lists adipocytes and smooth muscle cells as Not detected examples (HPA tissue IHC; general IHC practice). | Verify cell morphology and compare control staining. Treat the result as unresolved until its cellular pattern and background are accounted for (general IHC practice). |
| A section appears predominantly nuclear. | UniProt permits nuclear localisation during interphase, while HPA describes general tissue IHC as granular cytoplasmic (UniProt Q9NPJ3; HPA tissue IHC). | Check whether cytoplasmic granules are also present and whether nuclear color persists in the negative reagent control; avoid assigning specificity from compartment alone (general IHC practice). |
| IF/ICC shows junctional signal while tissue IHC looks cytoplasmic. | The assays have different reported observations: junctions in HPA ICC-IF and granular cytoplasm in HPA tissue IHC (HPA subcellular; HPA tissue IHC). | Interpret each image against its own assay and control evidence. Use the dedicated IF/ICC guide for that experiment; do not use the ICC-IF location as a tissue IHC scoring rule (HPA subcellular; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing ACOT13 staining
A07288-1 · IHCImmunohistochemistry of THEM2 in human liver tissue with THEM2 antibody at 2.5 μg/mL.
A07288-1 · IFImmunofluorescence of THEM2 in human liver tissue with THEM2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000112304-ACOT13/tissue/cerebral+cortex#imid_5316560
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000112304-ACOT13/tissue/colon#imid_5316494
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000112304-ACOT13/tissue/liver#imid_5316487
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000112304-ACOT13/tissue/kidney#imid_5316488
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019881; green = ACOT13, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019881; green = ACOT13, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACOT13 IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in hepatocytes; selected IHC caption: human liver). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected); on the liver slide, compare hepatocytes with adjacent non-hepatocyte cells for background, without assuming those cells are ACOT13-negative (HPA: High designation applies to hepatocytes).
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ACOT13 in A-431, U-251MG, with annotated localisation: Cell Junctions (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class; use ACOT13 knockout material, if available, as a biological specificity control (standard IHC practice). For chromogenic liver IHC, block endogenous peroxidase and assess pigment-related background alongside the controls (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07288-1 liver IHC caption does not state a fixative (selected IHC caption: fixative not stated). Retrieval dependency is unreported, so optimize antigen retrieval with the catalog antibody’s IHC-P protocol; the supplied evidence does not establish whether frozen sections or IF/ICC are easier. Liver pigment can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for ACOT13
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACOT13 IHC Tips
Troubleshoot ACOT13 staining in paraffin sections by checking retrieval, cell type, subcellular pattern and controls before interpreting chromogenic signal.
How should I retrieve ACOT13 in paraffin sections when the signal is weak?
Start with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval setting). If staining remains weak, compare a shorter heating time on adjacent sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Include a liver section and assess hepatocyte staining, since high expression in hepatocytes provides a reference for the run (HPA: High in hepatocytes). Record staining intensity and tissue integrity together: excessive retrieval can damage morphology or increase diffuse background (standard IHC practice). The selected antibody image documents liver staining at 2.5 µg/mL, but its caption does not state the fixative or retrieval method (A07288-1 caption).
Could fixation explain weak or uneven ACOT13 staining?
Target-specific fixation sensitivity for ACOT13 is unknown from the supplied evidence (ACOT13 evidence payload). Record fixation duration and processing conditions for each specimen, then compare similarly processed sections before changing retrieval or antibody concentration (standard IHC practice). Use adjacent sections to distinguish uneven preservation from a cell-specific staining pattern, and inspect folds, torn areas and poorly preserved regions before scoring (standard IHC practice). The selected liver image reports staining at 2.5 µg/mL but gives no fixative, so it cannot establish an optimal fixation condition (A07288-1 caption). HPA tissue patterns and the protein’s topology or acetylation sites do not establish fixation sensitivity (HPA tissue IHC; UniProt Q9NPJ3).
Which ACOT13 staining compartments should I accept in chromogenic IHC?
Expect a predominantly cytoplasmic, sometimes granular tissue pattern, while judging each stained cell against local morphology (HPA tissue IHC: general cytoplasmic expression with a granular pattern). Cytosolic, mitochondrial, nuclear and spindle localisation are annotated for ACOT13, with nuclear and cytoplasmic localisation during interphase and spindle localisation during mitosis (UniProt Q9NPJ3 subcellular location). Score convincing nuclear signal separately from cytoplasmic signal rather than combining them into one intensity value (standard IHC practice). Cell-junction localisation comes from cellular imaging and should be treated as a comparison point, not an assumed dominant pattern in paraffin tissue (HPA subcellular: Cell Junctions approved). Recheck diffuse staining that ignores cell boundaries against control sections (standard IHC practice).
Can this IHC pattern distinguish ACOT13 isoforms or an altered epitope?
Do not assign staining to one isoform without an antibody epitope map and isoform-specific validation, because 2 ACOT13 isoforms are listed (UniProt Q9NPJ3 isoforms). The record also lists chains beginning at residues 1 and 2 and several acetylated residues, including positions 1, 2, 27, 37 and 43 (UniProt Q9NPJ3 processing and modified residues). Those annotations identify possible epitope considerations but do not show that processing or acetylation changes this antibody’s tissue staining (UniProt Q9NPJ3; A07288-1 caption). If staining differs across specimens, compare matched sections and document the antibody’s stated immunogen before interpreting an epitope effect (standard IHC practice).
How can I cross-check ACOT13 localisation with multiplex IF?
Use the separate IF/ICC workflow to pair ACOT13 with an established marker for the cell type being examined, and include single-label controls for each channel (standard IF practice). For a liver comparison, identify hepatocytes before asking whether ACOT13 signal overlaps the expected cells (HPA: High in hepatocytes). Choose a spectrally separated fluorophore, favoring a far-red channel when tissue autofluorescence obscures shorter wavelengths, and check unstained tissue in each channel (standard IF practice). ACOT13 has no transmembrane segment and has intracellular localisation annotations, so use permeabilisation appropriate to the intracellular epitope and antibody instructions (UniProt Q9NPJ3 topology and subcellular location; standard IF practice). Do not transfer the liver IHC caption’s conditions to IF (A07288-1 caption).
What controls help separate ACOT13 signal from chromogenic background?
Run a no-primary control and inspect the same tissue regions for residual chromogen or endogenous enzyme activity (standard IHC practice). Include a peroxidase block when using peroxidase and DAB detection, and verify that the block reduces enzyme-derived staining without obscuring morphology (standard IHC practice). Compare high-expression hepatocytes or kidney tubular cells with a reported non-detected population, such as adipocytes, while accounting for differences in tissue processing (HPA: High in hepatocytes and kidney tubules; Not detected in adipocytes; standard IHC practice). If diffuse staining persists, titrate primary antibody and detection reagent on adjacent sections; the selected liver image reports 2.5 µg/mL for its staining, not a universal working concentration (A07288-1 caption).
How should I score heterogeneous ACOT13 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since the reported tissue pattern is generally cytoplasmic and granular (HPA tissue IHC: general cytoplasmic expression with a granular pattern). For each region, report the percentage of positive target cells and an intensity score, or calculate an H-score from the percentages at intensities 0–3 (standard IHC practice). If counting positive cells per mm², normalise to evaluable tissue area and report the number of target cells assessed (standard IHC practice). Keep hepatocyte and stromal measurements separate in liver sections, because hepatocytes are a reported high-expression population (HPA: High in hepatocytes). Apply one threshold across comparably processed sections and exclude folds, edges and necrosis from the scored area (standard IHC practice).
When is an apparent ACOT13-positive area likely to be artefactual?
Treat cytoplasmic granular staining in appropriately identified hepatocytes as more consistent with the tissue reference than staining limited to damaged section edges (HPA: High in hepatocytes; HPA tissue IHC: general cytoplasmic granular pattern; standard IHC practice). Reassess strong staining confined to necrosis, folds or cut edges, and compare it with the no-primary control (standard IHC practice). Apparent nuclear or spindle signal needs intact cell morphology and a compartment-specific check, because those locations are annotated but are not the general HPA tissue pattern (UniProt Q9NPJ3 subcellular location; HPA tissue IHC). Residual peroxidase staining after detection can mimic positivity, so check the peroxidase-blocked control before assigning such signal to ACOT13 (standard IHC practice).
Boster reagents
Best ACOT13 / Acyl-coenzyme A thioesterase 13 IHC Antibodies
A07288-1 has human liver IHC and IF images (catalog image captions) and lists human, mouse, and rat reactivity (catalog: reactivity).
Real IHC data
Anti-THEM2 ACOT13 Antibody
Cat # A07288-1
A07288-1 is listed for IHC-P and IF (catalog: applications). Its images show human liver IHC at 2.5 μg/mL and IF at 20 μg/mL (catalog image captions).
Which to pick: For paraffin-section tissue IHC, choose A07288-1: it lists IHC-P and shows human liver staining at 2.5 μg/mL; the caption does not report the fixative (catalog: applications; IHC image caption). For fluorescence, A07288-1 lists IF and shows human liver staining at 20 μg/mL; ICC validation is not listed (catalog: applications; IF image caption). For cross-species work, A07288-1 lists human, mouse, and rat reactivity and has a rabbit host, but the supplied images show human tissue only (catalog: reactivity, host, image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NPJ3 (ACO13_HUMAN, Acyl-coenzyme A thioesterase 13).
- Human Protein Atlas. ACOT13 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACOT13 subcellular location (ICC-IF): Mainly localized to the cell junctions..
- Human Protein Atlas. ACOT13 antibody validation summary (2 antibodies).
- Low expression of acyl-CoA thioesterase 13 is associated with poor prognosis in ovarian serous cystadenocarcinoma. Frontiers in genetics 2023 — PMC10323136.
- Characterization of the Lipid Metabolism in Bladder Cancer to Guide Clinical Therapy. Journal of oncology 2022 — PMC9484922.
- A novel nanotherapeutic strategy: rescuing nucleus pulposus cells from fatty acid metabolic disorder and pyroptosis through ACOT13 by Chinese herbal formula nanoparticles. Journal of nanobiotechnology 2026 — PMC12983563.
- PubMed PMID:10931946 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACOT13 (Acyl-coenzyme A thioesterase 13). Expected localisation Granular cytoplasm in tissue sections (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07288-1 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A07288-1); 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 Tris-EDTA buffer, pH 9.0 , 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-ACOT13, 2.5 μg/mL (datasheet A07288-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07288-1); Caveat: Antibody–RNA consistency is medium (HPA tissue IHC); Isoform / epitope: 2 isoforms; N-terminal processing may affect N-terminal epitopes (UniProt); Regulation: No regulatory stimulus specified (UniProt record). Evidence: UniProt Q9NPJ3, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cell junctions., Datasheet A07288-1, PMC12983563. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACOT13/immunohistochemistry-ihc-guide.