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ACAT1 / Acetyl-CoA acetyltransferase, mitochondrial · IHC design guide
Design Immunohistochemistry for ACAT1
Plan chromogenic ACAT1 IHC around the granular cytoplasmic tissue pattern (HPA tissue IHC). Use high-staining adrenal glandular cells as a reference (HPA tissue IHC) and start antibody A02008-1 at 2–5 μg/ml (datasheet A02008-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACAT1 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); mitochondrial protein (UniProt) | |
| Staining pattern | Most cell types show granular cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02008-1) | |
| Positive control | Adrenal gland+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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Smooth muscle cells show low staining (HPA tissue IHC) | |
| Regulation | Kidney/liver RNA enrichment (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; mature chain 34–427; epitope effect unknown (UniProt) |
Section 1
Recommended ACAT1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published paraffin-section methods (datasheet A02008-1; PMC11544177; PMC8994887; PMC6795108; PMC13614741). Check antibody identity before adapting them.
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02008-1) |
| Fixation | Image fixative and duration unreported (datasheet A02008-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A02008-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02008-1) |
| Primary antibody | Rabbit anti-ACAT1, 2-5 μg/ml (datasheet A02008-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02008-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02008-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ACAT1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A02008-1). The ovarian study explicitly used anti-SOAT1/ACAT1, so verify target identity before using that protocol (PMC8994887).
Section 2
What Is the Expected ACAT1 Staining Pattern?
ACAT1 is a mitochondrial protein with no annotated transmembrane segment (UniProt P24752 topology). In paraffin-section IHC, expect granular cytoplasmic staining, reported across many cell types (HPA: ubiquitous granular cytoplasmic profile). Strong examples include adrenal glandular cells, bronchial respiratory epithelial cells, and cerebellar Purkinje cells (HPA: High). HPA rates the tissue staining profile Enhanced, reflecting consistency with RNA expression data (HPA: Enhanced reliability).
| Granular cytoplasmic staining in adrenal glandular cells or bronchial respiratory epithelial cells. | This fits the reported ACAT1 tissue pattern and High staining in those cells (HPA: tissue IHC). Granularity is consistent with mitochondrial localization (UniProt P24752; HPA: subcellular). Assess the expected cells separately from adjacent tissue. |
| Predominantly nuclear, membranous, or extracellular staining. | These compartments conflict with the reported mitochondrial location and granular cytoplasmic IHC profile (UniProt P24752; HPA: tissue IHC and subcellular). Treat the pattern as suspect and check antibody specificity, detection background, and slide morphology before assigning it to ACAT1 (general IHC practice). |
| Strong staining in a cell population expected to stain weakly, such as smooth muscle cells. | HPA lists smooth muscle cells as Low (HPA: tissue IHC). Unexpectedly strong signal warrants comparison with nearby expected positive cells and a detection control; cross-reactivity or endogenous detection activity is possible, but this observation alone cannot identify the cause (general IHC practice). |
| Diffuse chromogen across tissue, empty spaces, or many unrelated compartments. | Field-wide haze obscures the reported granular cytoplasmic pattern (HPA: tissue IHC). Consider nonspecific reagent binding or endogenous detection activity, especially if a control omitting primary antibody also develops color (general chromogenic IHC practice). |
| No staining in adrenal glandular cells or bronchial respiratory epithelial cells. | These are reported High examples, so an absent signal calls for a run-level check (HPA: tissue IHC). Examine tissue preservation, antibody and detection steps, and a concurrent positive control (general IHC practice); HPA does not establish an ACAT1-specific fixation sensitivity. |
💡Expected ACAT1 appearanceCall a result consistent with ACAT1 when expected cells show distinct granular cytoplasmic chromogen, with strong signal possible in HPA High examples (HPA: tissue IHC; UniProt P24752: mitochondrion). Predominantly nuclear staining or uniform field-wide color is a suspect result requiring controls (general IHC practice).
| Tissue and cell selection | HPA reports High staining in several specific cell populations and Low staining in smooth muscle cells (HPA: tissue IHC). Use the named cell population when interpreting a section; an organ-level RNA designation does not establish that every cell in the organ stains strongly (HPA: tissue IHC and RNA specificity). |
| Antibody evidence | The tissue profile has Enhanced reliability, and 2 listed antibodies have Enhanced IHC validation (HPA: tissue IHC; HPA: HPA004428 and HPA007569). This supports the reported pattern but does not validate an unlisted antibody or make every unexpected stain specific. |
| Topology and processing | ACAT1 is mitochondrial, has no annotated transmembrane segment, and has a reported chain spanning residues 34–427 (UniProt P24752). These annotations support an intracellular interpretation. They do not identify the catalog antibody's epitope or predict its behavior after paraffin processing. |
| Isoforms | UniProt lists 2 isoforms (UniProt P24752). The supplied evidence gives no isoform-specific IHC pattern or antibody epitope, so differences between stained cells cannot be assigned to an isoform from this section alone. |
| IF/ICC: where should signal appear? | HPA places ACAT1 in mitochondria with enhanced subcellular evidence (HPA: ICC-IF). A punctate mitochondrial distribution is the relevant IF/ICC comparison; it does not replace the granular cytoplasmic reading of chromogenic tissue IHC (HPA: subcellular and tissue IHC). |
| Chromogenic detection activity | Endogenous enzyme activity can contribute color in chromogenic IHC (general IHC practice). A control omitting primary antibody helps distinguish detection-related color from antibody-dependent staining; no ACAT1-specific endogenous detection effect is established by the supplied sources. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported High cell population is blank. | The IHC run may lack adequate antibody or detection signal; tissue condition may also matter (general IHC practice). | Check a concurrent positive control, reagent sequence, antibody dilution, and detection system (general IHC practice). Use a named HPA High cell population for comparison (HPA: tissue IHC). Do not infer ACAT1-specific fixation sensitivity from the blank slide. |
| Color persists when primary antibody is omitted. | Primary-independent color can arise from endogenous detection activity or detection reagents (general chromogenic IHC practice). | Review the control and apply the detection system's appropriate endogenous-activity blocking step; then compare the corrected slide with expected granular cytoplasmic staining (general IHC practice; HPA: tissue IHC). |
| The entire section has diffuse background. | Nonspecific reagent binding, inadequate washing, or excess detection signal can obscure cell boundaries (general IHC practice). | Check blocking, wash steps, antibody dilution, and development time against the validated workflow (general IHC practice). Score ACAT1 only where a cellular granular pattern remains interpretable (HPA: tissue IHC). |
| Signal appears mainly in nuclei or along cell borders. | That distribution disagrees with mitochondrial localization and the granular cytoplasmic tissue profile (UniProt P24752; HPA: tissue IHC). | Review section morphology and background controls, then compare with a reported positive cell population or an independently validated antibody where available (general IHC practice; HPA: IHC validation). |
| Smooth muscle stains as strongly as nearby reported High cells. | HPA reports Low staining in smooth muscle cells; cross-reactivity or detection background is possible (HPA: tissue IHC; general IHC practice). | Confirm cell identity on the counterstained section and inspect a control omitting primary antibody (general IHC practice). Report the discrepancy rather than treating intensity alone as proof of ACAT1. |
| Kidney or liver staining seems inconsistent with an RNA-based expectation. | HPA calls ACAT1 RNA tissue enhanced in kidney and liver, but the supplied IHC examples do not establish a matching staining intensity for every cell there (HPA: RNA specificity and tissue IHC). | Interpret the observed cell type and compartment, then compare with the reported granular cytoplasmic profile and a named HPA High example (HPA: tissue IHC). Avoid assigning an unsupported cell-specific intensity. |
Experiment evidence
IHC & IF images showing ACAT1 staining
A02008-1 · IHCIHC analysis of ACAT1 using anti-ACAT1 antibody (A02008-1). ACAT1 was detected in a paraffin-embedded section of human liver cancer tissue.
A02008-1 · IHCIHC analysis of ACAT1 using anti-ACAT1 antibody (A02008-1). ACAT1 was detected in a paraffin-embedded section of human non-small cell lung c
A02008-1 · IFIF analysis of ACAT1 using anti-ACAT1 antibody (A02008-1). ACAT1 was detected in an immunocytochemical section of HELA cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000075239-ACAT1/tissue/cerebral+cortex#imid_1612128
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000075239-ACAT1/tissue/colon#imid_1612054
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000075239-ACAT1/tissue/liver#imid_1612063
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000075239-ACAT1/tissue/kidney#imid_1612064
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004428; green = ACAT1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004428; green = ACAT1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACAT1 IHC & IF
🧪Run bronchus first; respiratory epithelial cells should stain strongly (HPA: High in bronchus respiratory epithelial cells). HPA detects ACAT1 in all 45 scored tissues, so there is no supported negative tissue (HPA: no negative rows); use no-primary and isotype controls, and treat any weakly stained cells on the positive slide only as relative within-slide contrast, not confirmed ACAT1-negative cells.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: ACAT1 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 ACAT1 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Mitochondria (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a matched rabbit IgG isotype control (selected-SKU caption: rabbit primary), and ACAT1-knockout material as a biological specificity control. Quench endogenous peroxidase and inspect inflammatory cells for background in bronchus sections (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption). The reported IHC procedure uses heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody, but does not establish that retrieval is required (selected-SKU caption). Frozen sections cannot be judged easier from the supplied evidence; ICC-IF has a mitochondrial localization reference (HPA: mitochondria enhanced), while bronchial inflammatory cells may contribute endogenous peroxidase background (standard chromogenic IHC practice).
HPA tissue IHC evidence for ACAT1
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 → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ACAT1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ACAT1 IHC Tips
Use the catalog antibody’s paraffin section result as a starting point, then judge ACAT1 staining by its granular cytoplasmic pattern and mitochondrial localisation.
How should I retrieve ACAT1 in paraffin sections when staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02008-1). The selected image used this retrieval before 2 μg/ml primary antibody overnight at 4°C, so keep those conditions together for the first comparison (datasheet A02008-1). If staining remains weak, compare a shorter and a longer heating interval on adjacent sections while holding antibody concentration and DAB development constant; assess tissue integrity and granular cytoplasmic detail in each section (HPA: granular cytoplasmic expression). A different retrieval buffer is a fallback to test with controls, since no second buffer is documented for this SKU (datasheet A02008-1).
Could fixation account for variable ACAT1 staining across paraffin blocks?
Target specific sensitivity to fixation is unknown: the selected paraffin section caption does not state its fixative or fixation duration (datasheet A02008-1). Record each block’s fixative and time in fixative, then compare adjacent sections processed with the same EDTA pH 8.0 retrieval, 2 μg/ml primary antibody, and DAB development (datasheet A02008-1). Include a reference block in each run and check whether weak staining coincides with poor tissue morphology or uneven staining across the section. Do not attribute a difference to ACAT1 epitope damage until processing differences and run variation have been assessed; the caption supplies no fixation comparison (datasheet A02008-1).
What staining pattern should count as correctly localised ACAT1?
Expect granular cytoplasmic staining in chromogenic sections, consistent with mitochondrial ACAT1 rather than a continuous plasma membrane outline (HPA: ubiquitous granular cytoplasmic expression; UniProt P24752: mitochondrion, no transmembrane segment). Compare signal at a magnification that resolves cytoplasmic granules and keep the haematoxylin counterstain light enough to distinguish them from nuclei. The HPA subcellular assessment places ACAT1 in mitochondria, providing an independent compartment check for the tissue pattern (HPA: mitochondria, enhanced). If staining is chiefly nuclear, extracellular, or restricted to section edges, revisit background controls and retrieval before scoring it as ACAT1 (UniProt P24752: mitochondrion; HPA: granular cytoplasmic expression).
Can this antibody distinguish ACAT1 isoforms or modified epitopes in IHC?
ACAT1 has 2 annotated isoforms and a processed chain spanning residues 34–427 (UniProt P24752: isoforms and chain). Its record also lists acetyllysine and succinyllysine modifications, including alternative annotations at residues 66 and 174 (UniProt P24752: modified residues). The supplied IHC caption documents staining conditions but gives no epitope mapping or isoform discrimination for A02008-1, so interpret chromogenic intensity as antibody reactivity rather than an isoform specific readout (datasheet A02008-1). If isoform or modification status matters, seek mapped epitope information and an independently validated assay before assigning the tissue signal to either molecular form.
How can I check ACAT1 localisation by multiplex IF alongside this IHC result?
Treat IF as a separate assay: the supplied A02008-1 caption documents chromogenic staining of a paraffin section, with no IF fixation or antibody dilution specified (datasheet A02008-1). Multiplex ACAT1 with a marker for the cell population under study and, if needed, a mitochondrial marker; ask whether punctate cytoplasmic signal belongs to the marked cells and overlaps mitochondria (HPA: granular cytoplasmic expression; HPA: mitochondria, enhanced). Choose fluorophores after inspecting unstained tissue for autofluorescence and reserve a spectrally distinct channel for weak ACAT1 signal. For intracellular ACAT1, titrate permeabilisation that grants antibody access to the mitochondrial compartment while preserving organelle morphology (UniProt P24752: mitochondrial localisation, no transmembrane segment).
How do I separate ACAT1 staining from DAB or tissue background?
Start by inspecting a section without primary antibody, plus a section processed with the same 10% goat serum block, peroxidase conjugated secondary, and DAB workflow used in the selected image (datasheet A02008-1). A peroxidase block is a general chromogenic IHC step; assess its effectiveness where endogenous enzyme activity or blood could produce color, without treating that color as ACAT1. Titrate primary antibody and DAB development if diffuse brown signal obscures the expected granular cytoplasmic pattern (HPA: granular cytoplasmic expression). Compare suspect areas with intact tissue away from folds, edges, and necrosis, and retain identical development conditions across sections being compared.
What is a defensible way to quantify ACAT1 in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cells and regions before scoring, then record cytoplasmic staining intensity and the percentage of cells at each intensity to calculate an H score, or report percentage positive cells using a fixed threshold. If cell abundance is the endpoint, report positive cell density per mm² of viable tissue and normalise it to the relevant cell population or total evaluable cells. Keep retrieval at EDTA pH 8.0 and primary antibody at 2 μg/ml across comparison slides when following the selected image conditions (datasheet A02008-1). Exclude folds, necrosis, and edges using the same rules for every slide; score granular cytoplasmic signal rather than nonspecific diffuse color (HPA: granular cytoplasmic expression).
Which ACAT1 staining differences are biologically credible rather than artefacts?
A credible signal is granular and cytoplasmic, in keeping with mitochondrial localisation (HPA: granular cytoplasmic expression; UniProt P24752: mitochondrion). Cell context matters: HPA reports high staining in adrenal glandular cells and low staining in smooth muscle cells, so compare like cell populations before calling a tissue difference (HPA: adrenal glandular cells High; smooth muscle cells Low). Treat nuclear or extracellular color, edge accentuation, necrotic areas, and staining persisting without primary antibody as reasons to investigate artefact, including endogenous peroxidase in a DAB workflow. Recheck any proposed difference on intact regions processed under matched retrieval, antibody incubation, and chromogen conditions (datasheet A02008-1: EDTA pH 8.0, 2 μg/ml primary, DAB).
Boster reagents
Best ACAT1 / Acetyl-CoA acetyltransferase, mitochondrial IHC Antibodies
The IHC-validated antibody has paraffin-section images from three human tissues and an IF image from HeLa cells; human, mouse and rat reactivity is listed (catalog: A02008-1 images and reactivity).
Real IHC data
Anti-ACAT1 Antibody ®
Cat # A02008-1
The rendered SKU, A02008-1, has IHC images from paraffin sections of human liver cancer, non-small cell lung cancer and placenta (catalog: A02008-1 IHC captions). Its IF image shows HeLa cells (catalog: A02008-1 IF caption).
Which to pick: Choose A02008-1 for tissue IHC when a documented paraffin-section example is useful; its caption does not report the fixative (catalog: A02008-1 IHC caption). Choose A02008-1 for IF/ICC because both applications are listed and its IF image shows HeLa cells (catalog: A02008-1 applications and IF caption). For work across species, A02008-1 and the monoclonal M02008 both list human, mouse and rat reactivity, but the supplied IHC images document human tissue only for A02008-1 (catalog: reactivity, M02008 clone 29A55 and IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P24752 (THIL_HUMAN, Acetyl-CoA acetyltransferase, mitochondrial).
- Human Protein Atlas. ACAT1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACAT1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. ACAT1 antibody validation summary (2 antibodies).
- Target fishing and mechanistic insights of the natural anticancer drug candidate chlorogenic acid. Acta pharmaceutica Sinica. B 2024 — PMC11544177.
- Assessment of the diagnostic and prognostic relevance of ACAT1 and CE levels in plasma, peritoneal fluid and tumor tissue of epithelial ovarian cancer patients - a pilot study. BMC cancer 2022 — PMC8994887.
- ACAT1 and Metabolism-Related Pathways Are Essential for the Progression of Clear Cell Renal Cell Carcinoma (ccRCC), as Determined by Co-expression Network Analysis. Frontiers in oncology 2019 — PMC6795108.
- Cholesterol metabolism dysregulation in eyelid sebaceous gland carcinoma and its clinical significance. Indian journal of ophthalmology 2026 — PMC13614741.
- PubMed PMID:1979337 — UniProt-cited evidence.
- PubMed PMID:1684944 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACAT1 (Acetyl-CoA acetyltransferase, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02008-1). Validated IHC antibody: A02008-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02008-1); 2. Fixation: Image fixative and duration unreported (datasheet A02008-1); 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 retrieval: EDTA pH 8.0 (datasheet A02008-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02008-1); 8. Primary antibody: Rabbit anti-ACAT1, 2-5 μg/ml (datasheet A02008-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02008-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Smooth muscle cells show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; mature chain 34–427; epitope effect unknown (UniProt); Regulation: Kidney/liver RNA enrichment (HPA tissue RNA). Evidence: UniProt P24752, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A02008-1, PMC11544177, PMC8994887. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACAT1/immunohistochemistry-ihc-guide.