This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ACACB / Acetyl-CoA carboxylase 2 · IHC design guide
Design Immunohistochemistry for ACACB
This paraffin IHC guide uses adipocytes as a strong ACACB staining reference (HPA tissue IHC). It helps interpret observed cytoplasmic and nuclear staining (HPA tissue IHC) alongside the mitochondrial annotation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACACB 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 | Cytoplasmic and nuclear tissue staining (HPA tissue IHC); mitochondrial annotation (UniProt) | |
| Staining pattern | Strongest in adipocytes, with cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03668-2) | |
| Positive control | Adipose tissue+3 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Nuclear staining differs from the mitochondrial annotation (HPA tissue IHC; UniProt) | |
| Regulation | Adipose enriched expression (UniProt) | |
| Isoform / epitope | 3 isoforms; isoform specific epitope coverage is unknown (UniProt) |
Section 1
Recommended ACACB IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published ACACB chromogenic IHC protocol using mouse brain sections (PMC13125043).
| Sample | Paraffin-embedded rat testis tissue; fixative not specified (datasheet A03668-2) |
| Fixation | Image fixative and duration unreported (datasheet A03668-2); 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 A03668-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03668-2) |
| Primary antibody | Rabbit anti-ACACB, 0.5-1μg/ml (datasheet A03668-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03668-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03668-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ACACB-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues, most abundant in adipocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A03668-2); the published mouse brain excerpt does not specify retrieval (PMC13125043).
Section 2
What Is the Expected ACACB Staining Pattern?
ACACB is annotated as mitochondrial and has no transmembrane segment (UniProt O00763: location and topology). In tissue IHC, expect predominantly cellular staining in adipocytes, with medium staining in hepatocytes and breast adipocytes (HPA: tissue IHC). HPA also reports cytoplasmic and nuclear expression across several tissues; its tissue IHC reliability is Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Strong cellular staining in adipose tissue adipocytes; medium staining in liver hepatocytes or breast adipocytes (HPA: tissue IHC). | This follows the reported cell and intensity pattern. Assess staining in the named cells against adjacent tissue and controls; a positive tissue does not imply that every cell in the section should stain (HPA: tissue IHC; general IHC practice). |
| Cytoplasmic staining, with some nuclear signal in tissue sections (HPA: tissue IHC). | HPA reports both compartments, while UniProt assigns ACACB to mitochondria (HPA: tissue IHC; UniProt O00763: location). Nuclear staining alone therefore cannot confirm mitochondrial localization. Record the compartment and compare it with controls before interpreting an unusual pattern (general IHC practice). |
| Predominantly extracellular, surface-rim, or acellular deposit staining. | That distribution does not fit a mitochondrial protein without a transmembrane segment (UniProt O00763: location and topology). Treat it as a possible staining artefact and review morphology, detection controls, and the location of precipitate (general IHC practice). |
| Strong staining in adrenal glandular cells or bone marrow hematopoietic cells. | Those cells were reported as not detected in HPA tissue IHC (HPA: adrenal gland; bone marrow). Check for nonspecific antibody binding or endogenous detection activity before calling the signal ACACB; HPA's tissue observations do not prove every specimen will be negative (general IHC practice). |
| Diffuse color across cells, stroma, and blank spaces. | A widespread haze obscures the cell-specific pattern reported by HPA (HPA: tissue IHC). It may reflect background from detection reagents, incomplete blocking, or excessive antibody concentration; interpret only signal that remains distinguishable from negative controls (general IHC practice). |
💡Expected ACACB appearanceA convincing positive has high cellular staining in adipose adipocytes, with cytoplasmic signal expected from the mitochondrial annotation; dominant acellular deposits or strong staining in HPA-negative cell types warrants investigation (HPA: tissue IHC; UniProt O00763: location).
| Tissue and cell selection | Adipose adipocytes are a stronger positive reference than liver hepatocytes or breast adipocytes, which HPA scores medium; adrenal glandular cells are reported not detected (HPA: tissue IHC). Score the specified cells rather than whole-section color (general IHC practice). |
| Compartment evidence | UniProt assigns ACACB to mitochondria, yet HPA describes cytoplasmic and nuclear tissue staining (UniProt O00763: location; HPA: tissue IHC). Tissue IHC supports a cellular pattern but cannot by itself resolve subcellular localization (general IHC practice). |
| Isoforms and epitope coverage | UniProt lists three ACACB isoforms and reports isoform 3 protein in skeletal muscle, adipose tissue, and liver (UniProt O00763: isoforms and tissue specificity). The supplied evidence gives no antibody epitope, so isoform-specific recognition cannot be assumed. |
| Topology and processing | ACACB has no transmembrane segment, signal peptide, or propeptide in the supplied record (UniProt O00763: topology and processing). These annotations support an intracellular interpretation; they do not establish how paraffin processing affects antibody recognition. |
| Biotin-linked detection | ACACB has a biotinyl-binding domain and uses biotin in catalysis (UniProt O00763: domains and function). If an avidin–biotin detection system is used, assess endogenous biotin background with appropriate controls; that concern is general IHC practice, not evidence of ACACB-specific background. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in adipose adipocytes. | They are scored high by HPA, so a blank result conflicts with the reported tissue pattern (HPA: adipose tissue). Assay failure or specimen variation remains possible (general IHC practice). | Check tissue identity, positive and negative controls, antibody incubation, antigen retrieval, and chromogenic detection using the validated IHC workflow (general IHC practice). Do not infer ACACB absence from one blank section. |
| Adipocytes stain, but the entire section is dark. | The broad background reduces confidence in the cell-specific adipocyte pattern (HPA: tissue IHC). Incomplete blocking or overly strong detection can create diffuse color (general IHC practice). | Compare the no-primary control, inspect for precipitate, and adjust blocking, washing, or antibody concentration within the assay's validated range (general IHC practice). Reassess the named cells after background is controlled. |
| Strong color appears in HPA-negative cell types. | Adrenal glandular and bone marrow hematopoietic cells are reported not detected (HPA: tissue IHC). Nonspecific binding or endogenous detection activity is possible (general IHC practice). | Compare matched negative controls and morphology; if using avidin–biotin detection, check endogenous biotin controls (general IHC practice). Do not treat an unexpected positive as confirmed ACACB without independent support. |
| Staining appears only as extracellular deposits or tissue-edge color. | An acellular or edge-limited pattern does not match the intracellular localization annotation (UniProt O00763: location and topology). Deposited chromogen or section artefact is possible (general IHC practice). | Inspect the section and no-primary control, then repeat the validated stain if deposits obscure cellular interpretation (general IHC practice). Score only identifiable cells with interpretable morphology. |
| Nuclear staining seems stronger than cytoplasmic staining. | HPA reports nuclear as well as cytoplasmic tissue expression, whereas UniProt annotates mitochondrial localization (HPA: tissue IHC; UniProt O00763: location). Neither source establishes that nuclear dominance is specific. | Document the compartment and compare controls and other positive cells. Treat the localization as unresolved when nuclear staining is dominant; avoid assigning a mechanism from this IHC result alone (general IHC practice). |
| Can IF confirm the IHC compartment pattern? | HPA supplies no ICC-IF images or main subcellular location for ACACB (HPA: subcellular record). Its IHC validation status does not establish an IF staining pattern (HPA: antibody record). | Use the separate IF/ICC guide for that application. Interpret this page's cell and intensity expectations as tissue IHC evidence, and do not transfer its validation status to IF (HPA: tissue IHC; antibody record). |
Experiment evidence
IHC & IF images showing ACACB staining
A03668-2 · IHCIHC analysis of ACACB using anti-ACACB antibody (A03668-2). ACACB was detected in paraffin-embedded section of rat testis tissue. Heat media
M03668-1 · IHCHuman breast carcinoma was stained with anti-ACACB rabbit antibody
M03668-1 · IHCHuman kidney was stained with anti-ACACB rabbit antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000076555-ACACB/tissue/cerebral+cortex#imid_19908162
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000076555-ACACB/tissue/colon#imid_19907888
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000076555-ACACB/tissue/liver#imid_19907881
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000076555-ACACB/tissue/kidney#imid_19907882
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACACB IHC & IF
🧪Run adipose tissue first; adipocytes should stain strongly (HPA: High in adipocytes). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected), and expect nonadipocyte cells on the adipose slide to remain at background unless independently validated (HPA: High is assigned to adipocytes).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ACACB; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) slide, a species- and class-matched nonimmune rabbit IgG isotype control, and ACACB-knockout tissue as a biological negative (A03668-2 caption: rabbit primary; standard IHC practice). For HRP/DAB staining, quench endogenous peroxidase; for IF on adipose tissue, check tissue autofluorescence in a no-primary control (A03668-2 caption: HRP/DAB; standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state the fixative (A03668-2 caption). Heat-mediated retrieval in EDTA at pH 8.0 is documented for the catalog antibody in a paraffin section; comparative ease of frozen-section staining or IF is unreported (A03668-2 caption). In adipose sections, extracted lipid can leave clear adipocyte vacuoles that complicate localisation (standard histology practice).
HPA tissue IHC evidence for ACACB
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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACACB IHC Tips
Use the catalog antibody’s paraffin-section IHC conditions as the starting point, then evaluate staining against ACACB’s reported tissue distribution and localisation.
Which retrieval conditions should I start with for ACACB in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA buffer at pH 8.0 (datasheet A03668-2). The catalog antibody’s rat testis image used paraffin sections, followed by 10% goat serum blocking and 1 μg/ml primary antibody overnight at 4°C (caption A03668-2). Keep retrieval conditions consistent across the study before comparing staining intensity between specimens. If signal is weak, adjust heating time incrementally while checking tissue morphology and a positive control. Record the actual heating time and temperature for each run; the supplied caption specifies the buffer and pH but gives no retrieval duration (caption A03668-2).
How should I assess whether fixation is reducing ACACB staining?
The supplied ACACB paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (caption A03668-2). Record the fixative, fixation duration, tissue thickness and processing schedule for each specimen, then compare sections processed alike using the same retrieval and detection conditions. Include a known positive compartment, such as adipocytes, alongside the study sections (HPA: High in adipocytes). If staining differs between processing batches, repeat the comparison with matched sections before attributing the change to ACACB expression. The caption’s 1 μg/ml overnight primary incubation documents staining conditions, but cannot establish which fixation conditions preserve its epitope (caption A03668-2).
What staining pattern is plausible for ACACB in tissue IHC?
ACACB is annotated to mitochondria and has no transmembrane segment, so examine cytoplasmic staining in the context of mitochondrial distribution (UniProt O00763 localisation and topology). HPA reports cytoplasmic and nuclear expression across several tissues, with the strongest reported tissue staining in adipocytes (HPA tissue IHC). Treat nuclear-only staining cautiously because the two sources give different localisation descriptions; inspect the corresponding cytoplasm and an appropriate positive compartment. In testis, HPA reports high staining in elongated or late spermatids, while the catalog image demonstrates staining in a rat testis paraffin section (HPA tissue IHC; caption A03668-2). Compare cell types within intact tissue rather than interpreting colour alone.
Could isoforms or epitope accessibility explain uneven ACACB staining?
ACACB has 3 reported isoforms, and isoform 3 has protein-level evidence in skeletal muscle, adipose tissue and liver (UniProt O00763 isoforms and tissue specificity). The supplied antibody caption does not identify its epitope or establish which isoforms it detects (caption A03668-2). ACACB also contains annotated phosphorylation sites, including serines 35, 91 and 95, but their effects on this antibody’s binding are unknown (UniProt O00763 modified residues). Compare tissues using identical retrieval and antibody conditions, then seek epitope or isoform validation before assigning a staining difference to splicing or phosphorylation. Avoid treating loss of signal after harsher retrieval as evidence for a particular isoform.
How can I assess ACACB by multiplex IF alongside the IHC guide?
Treat multiplex IF as a separate validation experiment: the supplied product evidence documents paraffin-section chromogenic IHC, not IF performance (caption A03668-2). In adipose tissue, pair ACACB with an adipocyte marker such as perilipin to assess the cell type expected to stain strongly (HPA: High in adipocytes). Choose spectrally separated fluorophores, favour a far-red ACACB channel when tissue autofluorescence is troublesome, and include single-stain and unstained controls. Because ACACB has no transmembrane segment and is annotated to mitochondria, test gentle permeabilisation after fixation so intracellular epitopes are accessible (UniProt O00763 topology and localisation). Confirm that permeabilisation preserves cell morphology and marker separation before comparing signal across specimens.
How do I reduce diffuse or misleading DAB background?
Check background against a no-primary control and inspect areas with intact morphology before changing the ACACB antibody concentration. The catalog image used 10% goat serum blocking, 1 μg/ml primary antibody and peroxidase-based DAB detection (caption A03668-2). Include an endogenous peroxidase block as a general chromogenic IHC step, and keep DAB development time consistent across comparison slides. If using a biotin-based detection system, assess endogenous biotin background with an appropriate control; ACACB is a biotin-dependent enzyme, but that fact alone does not establish background in these sections (UniProt O00763 function). Persistent diffuse staining calls for reassessing blocking, washes and detection chemistry before interpreting cell-specific signal.
How should I score ACACB staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before examining outcomes; HPA reports high ACACB staining in adipocytes and in elongated or late spermatids (HPA tissue IHC). For chromogenic sections, report the percentage of positive target cells and an H-score using intensity classes 0–3, giving a possible range of 0–300. Alternatively, report positive-cell density per mm² when cell abundance varies between fields. Normalise stained-cell counts to all eligible cells of the same type, or stained area to the measured viable tissue area, and exclude folds or necrotic regions. Use identical retrieval, detection and scoring thresholds across specimens, with blinded review where feasible.
How can I distinguish genuine ACACB signal from tissue artefact?
Prioritise staining in plausible cells and compartments: adipocytes and elongated or late spermatids are reported as strongly positive, while ACACB is annotated to mitochondria (HPA tissue IHC; UniProt O00763 localisation). HPA also describes cytoplasmic and nuclear staining, so a nuclear component needs evaluation alongside cytoplasmic signal rather than automatic rejection (HPA tissue IHC). Compare suspected positives with no-primary controls and with nearby intact cells under the same DAB development conditions. Signal confined to section edges, folds or necrotic tissue suggests processing artefact; staining that persists without primary antibody suggests background from detection chemistry or endogenous peroxidase. Resolve discrepancies with matched processing and an independent antibody or orthogonal expression evidence before assigning biological meaning.
Boster reagents
Best ACACB / Acetyl-CoA carboxylase 2 IHC Antibodies
Two anti-ACACB antibodies have IHC images from rat and human tissues (catalog IHC captions). Both list human, mouse, and rat reactivity; one also lists IF (catalog applications/reactivity).
Real IHC data
Anti-Acetyl Coenzyme A Carboxylase/ACACB Antibody ®
Cat # A03668-2
Real IHC data
Anti-ACACB Rabbit Monoclonal Antibody
Cat # M03668-1
A03668-2 lists IHC and shows staining in a rat paraffin-embedded testis section (A03668-2 applications and IHC caption). M03668-1 lists IHC and IF and shows IHC staining in human breast carcinoma and kidney (M03668-1 applications and IHC captions).
Which to pick: For rat paraffin-section IHC, choose A03668-2: its caption reports EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody (A03668-2 IHC caption). For human tissue IHC or IF/ICC development, choose the monoclonal M03668-1: it has human tissue IHC images and lists IF, although no IF image or ICC validation is supplied (M03668-1 description, applications and image captions). Both list human, mouse, and rat reactivity; the fixative is unreported in their IHC captions (catalog reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00763 (ACACB_HUMAN, Acetyl-CoA carboxylase 2).
- Human Protein Atlas. ACACB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACACB subcellular location (ICC-IF): Highest expression in MCF-7: 16.2 nTPM.
- Human Protein Atlas. ACACB antibody validation summary (1 antibodies).
- ACACB is a novel metabolism-related biomarker in the prediction of response to cetuximab therapy inmetastatic colorectal cancer. Acta biochimica et biophysica Sinica 2022 — PMC9828296.
- Oleic acid improves pathological changes in Aβ1-42-induced astrocytes and Alzheimer's disease mouse models through PKA/ACACB/CPT1A. Frontiers in neuroscience 2026 — PMC13125043.
- Acetyl Co-A Carboxylase Inhibition Halts Hyperglycemia Induced Upregulation of De Novo Lipogenesis in Podocytes and Proximal Tubular Cells. Metabolites 2022 — PMC9610518.
- PubMed PMID:9099716 — UniProt-cited evidence.
- PubMed PMID:16854592 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACACB (Acetyl-CoA carboxylase 2). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC); mitochondrial annotation (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03668-2). Validated IHC antibody: A03668-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat testis tissue; fixative not specified (datasheet A03668-2); 2. Fixation: Image fixative and duration unreported (datasheet A03668-2); 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 A03668-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03668-2); 8. Primary antibody: Rabbit anti-ACACB, 0.5-1μg/ml (datasheet A03668-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03668-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Nuclear staining differs from the mitochondrial annotation (HPA tissue IHC; UniProt); Isoform / epitope: 3 isoforms ; isoform specific epitope coverage is unknown (UniProt); Regulation: Adipose enriched expression (UniProt). Evidence: UniProt O00763, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in MCF-7: 16.2 nTPM, Datasheet A03668-2, PMC13125043. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACACB/immunohistochemistry-ihc-guide.