This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ACADSB / Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial · IHC design guide
Design Immunohistochemistry for ACADSB
Plan chromogenic IHC of ACADSB in paraffin sections using hepatocytes as a high-staining reference (HPA tissue IHC). The catalog antibody is listed at 2–5 μg/mL for human IHC-P (datasheet A08652-1), and granular cytoplasmic staining is reported in most tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACADSB 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 matrix (UniProt) | |
| Staining pattern | Granular cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08652-1) | |
| Positive control | Breast+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 A08652-1) | |
| Caveat | Tissue staining has medium agreement with RNA data (HPA tissue IHC) | |
| Regulation | Liver-enhanced RNA expression (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; mature chain aa 34–432 (UniProt) |
Section 1
Recommended ACADSB IHC & IF Protocols
The catalog antibody’s IHC protocol (datasheet A08652-1) is accompanied by three published ACADSB IHC methods (PMC6506390; PMC8791850; PMC12585056).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A08652-1) |
| Fixation | Image fixative and duration unreported (datasheet A08652-1); 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 A08652-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08652-1) |
| Primary antibody | Rabbit anti-ACADSB, 2-5μg/ml (datasheet A08652-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08652-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08652-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ACADSB-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A08652-1); citrate retrieval is a published alternative (PMC6506390).
Section 2
What Is the Expected ACADSB Staining Pattern?
ACADSB is a mitochondrial matrix protein with no transmembrane segment (UniProt P45954 topology). In paraffin sections, expect granular cytoplasmic staining, particularly in hepatocytes, kidney distal tubules, and several glandular cell populations (HPA: tissue IHC). HPA rates the tissue profile Enhanced, with medium agreement between staining and RNA data; external verification remains pending (HPA: reliability).
| Granular cytoplasmic staining in hepatocytes, kidney distal tubules, or listed glandular cells (HPA: High). | This fits the reported tissue pattern and mitochondrial matrix location (HPA: tissue IHC; UniProt P45954). Judge intensity within the identified cell population: HPA reports high staining for these cells, while its broader description is granular cytoplasmic expression in most tissues (HPA: tissue IHC). |
| Predominantly nuclear, membranous, or extracellular staining, without convincing cytoplasmic granules. | Treat this as a localization mismatch that needs investigation, rather than assigning it to ACADSB: the protein is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt P45954 topology). Compare morphology and controls before interpreting an unusual compartment as specific staining (general IHC practice). |
| Strong staining in adipocytes, lung alveolar cells, or esophageal squamous epithelial cells (HPA: Not detected). | These cell populations are reported as not detected, so strong signal warrants a check for cross-reactivity or endogenous chromogen activity (HPA: tissue IHC; general IHC practice). A mismatch alone does not identify the cause; verify the cell type and inspect detection controls (general IHC practice). |
| Uniform haze across cells and surrounding tissue, with little cell-specific granular detail. | Diffuse background obscures the expected granular cytoplasmic pattern (HPA: tissue IHC). Check the no-primary control, blocking, washing, and chromogen development as general IHC troubleshooting steps; this appearance does not establish an ACADSB-specific fixation or epitope effect (general IHC practice). |
| No convincing signal in hepatocytes or kidney distal tubules despite intact tissue morphology. | Both are reported as high-staining populations, making them useful checks for an apparently negative run (HPA: tissue IHC). Review antibody identity and the recorded retrieval and detection steps before calling the sample negative; this record supplies no target-specific fixation sensitivity or retrieval condition (general IHC practice). |
💡Expected ACADSB appearanceCall a result consistent when hepatocytes, kidney distal tubules, or listed glandular cells show relatively strong granular cytoplasmic staining (HPA: High; HPA: tissue IHC); dominant nuclear, membranous, or extracellular signal is a localization mismatch (UniProt P45954 topology).
| Compartment and cellular distribution | ACADSB is assigned to the mitochondrial matrix, while tissue IHC describes granular cytoplasmic expression (UniProt P45954; HPA: tissue IHC). Read chromogenic granules in their tissue context; light microscopy does not by itself prove that each granule is mitochondrial (general IHC practice). |
| Choice of comparison tissue | HPA reports high staining in hepatocytes, kidney distal tubules, and specified glandular cells, but not-detected staining in several other named populations (HPA: tissue IHC). Compare the same cell type when judging a result; a whole-section average can conceal cell-specific differences (general IHC practice). |
| Strength of the reference pattern | HPA lists the tissue profile as Enhanced and both listed antibodies as IHC Enhanced, while describing only medium staining–RNA consistency and pending external verification (HPA: reliability; HPA: antibodies). Use the profile as a reference pattern, with its stated verification limit. |
| Protein processing and isoforms | UniProt lists a processed chain spanning residues 34–432 and two isoforms (UniProt P45954). The supplied record gives no antibody epitope or isoform-specific IHC pattern, so it cannot predict which form produces staining or explain a particular negative section. |
| IF/ICC Q: Where should signal localize? | A: HPA reports supported mitochondrial localization in ICC-IF and lists HPA041458 as ICC Supported (HPA: subcellular; HPA: antibodies). This is a localization reference for the separate IF/ICC guide, with no IF/ICC protocol specified here. |
| Situation | Likely cause | Next action |
|---|---|---|
| A hepatocyte or kidney distal-tubule control is blank. | The result conflicts with HPA's high staining in those cell populations; the record does not establish why (HPA: tissue IHC). | Confirm the cell type and antibody identity, then check the documented retrieval, primary-antibody, detection, and chromogen steps against the run record (general IHC practice). Do not infer ACADSB-specific fixation sensitivity. |
| The slide has widespread brown haze. | Background may arise from nonspecific binding, endogenous detection activity, or overdevelopment (general IHC practice). | Inspect a no-primary control, blocking and wash steps, and chromogen development; repeat with corrected general IHC conditions if needed (general IHC practice). Look for recovery of the granular cell pattern (HPA: tissue IHC). |
| Signal is mainly nuclear or outlines cell membranes. | That distribution conflicts with the mitochondrial matrix assignment and granular cytoplasmic tissue profile (UniProt P45954; HPA: tissue IHC). | Check tissue morphology and detection controls, and compare the suspect cells with a high-staining reference cell population (HPA: High; general IHC practice). Treat the compartment mismatch as unresolved until corroborated. |
| A reported not-detected cell population stains strongly. | Cross-reactivity or endogenous chromogen activity is possible; staining alone cannot distinguish them (HPA: tissue IHC; general IHC practice). | Verify the cell identity and inspect no-primary and detection controls (general IHC practice). Compare with a reported high-staining population on a suitable section before interpreting the unexpected signal (HPA: tissue IHC). |
| A section looks weak overall despite some positive cells. | Cell populations differ in the HPA profile: some are high, some low, and some not detected (HPA: tissue IHC). | Score identified cell populations separately, then compare like cells across sections (general IHC practice). Use the HPA levels as a reference and avoid treating low staining as proof of a failed run (HPA: tissue IHC). |
| Two antibodies give different tissue patterns. | Both listed antibodies have IHC Enhanced status, but the tissue profile still has medium staining–RNA consistency and awaits external verification (HPA: antibodies; HPA: reliability). | Document each antibody and compare the same cell populations and controls across sections (general IHC practice). Report the disagreement rather than assigning an unsupported epitope, isoform, or fixation mechanism. |
Experiment evidence
IHC & IF images showing ACADSB staining
A08652-1 · IHCIHC analysis of ACADSB using anti-ACADSB antibody (A08652-1). ACADSB was detected in a paraffin-embedded section of human liver cancer tissu
A08652-1 · IHCIHC analysis of ACADSB using anti-ACADSB antibody (A08652-1). ACADSB was detected in a paraffin-embedded section of human liver tissue. Heat
A08652-1 · IFIF analysis of SBCAD/ACADSB using anti-SBCAD/ACADSB antibody (A08652-1). SBCAD/ACADSB was detected in immunocytochemical section of Mcf-7 ce
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196177-ACADSB/tissue/cerebral+cortex#imid_20957439
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196177-ACADSB/tissue/colon#imid_20957373
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196177-ACADSB/tissue/liver#imid_20957374
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000196177-ACADSB/tissue/kidney#imid_20957375
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041458; green = ACADSB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041458; green = ACADSB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACADSB IHC & IF
🧪Run kidney first: distal tubules should stain strongly (HPA: High in kidney distal tubules). Use bone marrow hematopoietic cells as a negative tissue (HPA: Not detected in bone marrow hematopoietic cells); on the kidney slide, cells outside distal tubules should show only background unless independently shown to express ACADSB (HPA: High in distal tubules).
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ACADSB in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a concentration-matched nonimmune rabbit IgG control matched to the primary antibody’s clonality, and ACADSB knockout material as a biological negative (caption: rabbit primary antibody; standard IHC controls). Block endogenous peroxidase and check kidney background before interpreting DAB signal (caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: The selected IHC caption reports heat retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml overnight at 4°C, but reports no fixative or target-specific fixation window or effect (caption: A08652-1). It does not establish that retrieval is required, or that frozen sections are easier (caption: paraffin section with heat retrieval). For IF/ICC, mitochondrial localisation is supported, but ease relative to IHC is unreported; assess kidney tubular background when interpreting staining (HPA: Mitochondria supported; standard IHC practice).
HPA tissue IHC evidence for ACADSB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Distal tubules | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | 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 → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACADSB IHC Tips
Troubleshoot ACADSB staining in paraffin sections using the catalog antibody’s tissue IHC conditions, mitochondrial localisation, and cell specific reference patterns.
Which antigen retrieval should I start with for ACADSB in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A08652-1). The selected image used this retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A08652-1). If staining is weak, optimise heating and cooling while keeping the buffer and primary conditions recorded, then compare sections processed together (standard IHC practice). Judge improvement by clearer granular cytoplasmic signal in hepatocytes, rather than by an overall rise in DAB, because hepatocytes are reported as high and the usual tissue pattern is granular cytoplasmic (HPA: liver hepatocytes High; HPA: granular cytoplasmic expression).
How should I troubleshoot fixation when ACADSB staining is weak or uneven?
The selected paraffin section caption does not state a fixative, so target specific fixation sensitivity is unknown (datasheet A08652-1: fixative not stated). Record the fixative, fixation interval, section thickness, and time before processing for each specimen so that cases can be compared consistently (standard IHC practice). Run a well preserved reference section alongside each batch, using the documented EDTA pH 8.0 retrieval and 2 μg/ml overnight primary incubation as a starting point (datasheet A08652-1). If a poorly preserved region stains differently, compare intact cells within the same section before attributing the difference to ACADSB abundance (standard IHC practice).
Where should convincing ACADSB staining appear in tissue cells?
Expect granular cytoplasmic staining, consistent with a protein assigned to the mitochondrial matrix and with the reported tissue IHC profile (UniProt P45954: mitochondrial matrix; HPA: granular cytoplasmic expression). In liver, assess hepatocytes specifically, since they are reported as high for ACADSB (HPA: liver hepatocytes High). A predominantly nuclear, sharply membranous, or diffuse extracellular DAB pattern conflicts with that localisation and calls for review of morphology and controls (UniProt P45954: mitochondrial matrix; standard IHC practice). Because ACADSB has no annotated transmembrane segment, do not interpret a crisp cell surface outline alone as convincing target localisation (UniProt P45954: no transmembrane segment).
Could isoforms or epitope processing explain discordant ACADSB staining?
ACADSB has 2 listed isoforms, but the supplied evidence does not map this antibody’s epitope to either one (UniProt P45954: isoforms 1 and 2; datasheet A08652-1: epitope not stated). The annotated protein chain spans residues 34–432, so an epitope assignment would be needed before claiming that processing changes antibody recognition (UniProt P45954: chain 34–432). Acetylation or succinylation is annotated at residue 70 and phosphorylation at 183, without evidence here that these modifications alter this antibody’s IHC signal (UniProt P45954: modified residues; datasheet A08652-1: epitope not stated). Compare staining with cell morphology and a consistent retrieval batch before proposing an isoform or modification effect (standard IHC practice).
How can I check an ACADSB IHC pattern with multiplex immunofluorescence?
For a parallel IF check, pair ACADSB with a marker identifying the expected cell population, such as hepatocytes in liver, and assess whether its signal occupies the same cells (HPA: liver hepatocytes High; standard IF practice). Include a mitochondrial marker when testing compartment agreement, since ACADSB is assigned to the mitochondrial matrix (UniProt P45954: mitochondrial matrix; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained or single colour control to distinguish signal from tissue autofluorescence (standard IF practice). Because the target lies within mitochondria, optimise cell and organelle membrane access for the antibody’s unknown epitope while monitoring morphology; do not transfer the paraffin IHC conditions directly to IF (UniProt P45954: mitochondrial matrix; datasheet A08652-1: epitope not stated; standard IF practice).
How do I reduce widespread brown background without losing ACADSB signal?
First inspect a no primary control and apply a peroxidase block before DAB development to reveal endogenous enzyme activity or detection background (standard IHC practice). The selected paraffin section used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase conjugated secondary before DAB (datasheet A08652-1). If background persists, compare antibody titrations and washing conditions on adjacent sections while holding retrieval at EDTA pH 8.0 (datasheet A08652-1; standard IHC practice). Preserve granular cytoplasmic staining in an expected positive population while judging background in cell free spaces and the no primary control (HPA: granular cytoplasmic expression; standard IHC practice).
What scoring method makes ACADSB IHC comparisons interpretable? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring: hepatocytes are reported as high, whereas adipocytes are reported as not detected (HPA: liver hepatocytes High; HPA: adipose adipocytes Not detected). Within a fixed region of viable tissue, record the percentage of positive target cells and an intensity based H-score, using the same thresholds and DAB exposure across batches (standard IHC quantification practice). Normalise positive counts to the number of eligible target cells, or positive area to the analysed viable area in mm², rather than to the whole slide (standard IHC quantification practice). Report batch controls and separate granular cytoplasmic signal from diffuse pigment or edge staining during scoring (HPA: granular cytoplasmic expression; standard IHC practice).
When should I doubt an apparent ACADSB positive result?
Treat granular cytoplasmic staining in expected cells as more persuasive than nuclear, cell surface, or extracellular colour, because ACADSB is mitochondrial and the reported tissue pattern is granular cytoplasmic (UniProt P45954: mitochondrial matrix; HPA: granular cytoplasmic expression). Check cell identity: hepatocytes are reported as high, while adipocytes and bone marrow hematopoietic cells are reported as not detected (HPA: liver hepatocytes High; HPA: adipose adipocytes Not detected; HPA: bone marrow hematopoietic cells Not detected). Discount colour concentrated at section edges or in necrotic areas, and investigate signal in a no primary control or after inadequate peroxidase blocking (standard IHC practice). Interpret apparent exceptions cautiously because the tissue IHC evidence has medium agreement with RNA data and awaits external verification (HPA: Enhanced reliability description).
Boster reagents
Best ACADSB / Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial IHC Antibodies
A08652-1 has IHC images from human paraffin liver and liver cancer sections and an IF image from Mcf-7 cells (A08652-1 image captions).
Real IHC data
Anti-SBCAD/ACADSB Antibody ®
Cat # A08652-1
A08652-1 is listed for IHC and IF/ICC (A08652-1 catalog applications). Its IHC images show human paraffin liver and liver cancer sections, while its IF image shows Mcf-7 cells (A08652-1 image captions).
Which to pick: For tissue IHC, choose A08652-1: its own images show staining in human paraffin sections, with 2 μg/ml primary antibody after heat retrieval in EDTA at pH 8.0 (A08652-1 IHC image captions). For IF/ICC, the same SKU has an image from Mcf-7 cells; its listed IHC/IF reactivity includes human, mouse, rat and monkey, though the supplied IHC images demonstrate human tissue only (A08652-1 IF image caption; A08652-1 catalog reactivity; A08652-1 IHC image captions). The IHC captions identify paraffin sections but do not report the fixative (A08652-1 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P45954 (ACDSB_HUMAN, Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial).
- Human Protein Atlas. ACADSB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACADSB subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. ACADSB antibody validation summary (2 antibodies).
- Lipid metabolism-related gene signature predicts prognosis and depicts tumor microenvironment immune landscape in gliomas. Frontiers in immunology 2023 — PMC9968762.
- Loss of BCAA Catabolism during Carcinogenesis Enhances mTORC1 Activity and Promotes Tumor Development and Progression. Cell metabolism 2019 — PMC6506390.
- Decreased Expression of ACADSB Predicts Poor Prognosis in Clear Cell Renal Cell Carcinoma. Frontiers in oncology 2021 — PMC8791850.
- Unveiling and verification of mitochondria-related genes as potential diagnostic biomarkers in ulcerative colitis based on bioinformatics analysis and experimental validation. PloS one 2025 — PMC12585056.
- PubMed PMID:7698750 — UniProt-cited evidence.
- PubMed PMID:11013134 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACADSB (Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08652-1). Validated IHC antibody: A08652-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A08652-1); 2. Fixation: Image fixative and duration unreported (datasheet A08652-1); 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 A08652-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08652-1); 8. Primary antibody: Rabbit anti-ACADSB, 2-5μg/ml (datasheet A08652-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08652-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08652-1); Caveat: Tissue staining has medium agreement with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; mature chain aa 34–432 (UniProt); Regulation: Liver-enhanced RNA expression (HPA tissue RNA). Evidence: UniProt P45954, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A08652-1, PMC6506390, PMC8791850. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACADSB/immunohistochemistry-ihc-guide.