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ACADL / Long-chain specific acyl-CoA dehydrogenase, mitochondrial · IHC design guide
Design Immunohistochemistry for ACADL
Plan chromogenic ACADL IHC in paraffin sections with the catalog antibody's 1:50–1:200 dilution range (datasheet: IHC). Compare high staining in thyroid glandular cells and type II alveolar cells with undetected adipocytes, and score the cytoplasmic mitochondrial pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACADL 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 staining with a mitochondrial pattern (HPA tissue IHC) | |
| Staining pattern | Mitochondrial-pattern cytoplasm in thyroid glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Cell mix affects intensity: type II alveolar cells stain high (HPA tissue IHC) | |
| Regulation | Tissue and cell type dependent abundance (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms; mature chain spans 31–430 (UniProt) |
Section 1
Recommended ACADL IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published ACADL IHC methods for chordoid meningioma (PMC13081255) and human hepatocellular carcinoma (PMC7096519).
| Sample | FFPE rat ovary formalin fixed paraffin embedded tissue (datasheet A07206) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A07206); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-ACADL, 1:50-1:200 (datasheet A07206) |
| 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 | ACADL-positive staining in collecting ducts of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with a mitochondrial pattern in several tissues, most abundant in thyroid gland. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule).
Section 2
What Is the Expected ACADL Staining Pattern?
ACADL is a mitochondrial matrix protein with no transmembrane segment (UniProt P28330: location and topology). In paraffin section IHC, expect cytoplasmic staining with a mitochondrial pattern, especially in thyroid glandular cells, kidney collecting ducts and lung alveolar type II cells (HPA: tissue IHC, High in each cell type). HPA rates the tissue staining Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Granular cytoplasmic staining in thyroid glandular cells, kidney collecting ducts or alveolar type II cells. | This fits a mitochondrial matrix target and HPA's reported mitochondrial pattern; each listed cell type stains High (UniProt P28330: location; HPA: tissue IHC). Compare cells within the section, because a positive tissue does not imply uniform staining across every cell (general IHC interpretation). |
| Signal is predominantly nuclear, confined to the cell surface or deposited outside cells. | Those compartments conflict with the reported mitochondrial matrix location and lack of a transmembrane segment (UniProt P28330: location and topology). Review morphology and controls before calling the signal ACADL; chromogen precipitate or nonspecific detection can mislead compartment assignment (general IHC practice). |
| Strong staining appears in adipocytes or bone marrow hematopoietic cells. | HPA reports ACADL as Not detected in those specific cell types (HPA: tissue IHC). Check cell identification and background; cross-reactivity or endogenous detection activity are possibilities, but discordance alone does not establish either cause (general IHC interpretation). |
| Broad, hazy chromogen obscures cell borders and the expected cytoplasmic pattern. | This cannot be scored as a convincing mitochondrial pattern (HPA: tissue IHC profile; general IHC interpretation). Inspect the negative detection control and compare background across the section; nonspecific binding or detection background can produce haze (general IHC practice). |
| No signal is visible in an otherwise interpretable lung type II, thyroid glandular or kidney collecting duct compartment. | These are High HPA IHC reference cell types, so their absence prompts a technical review (HPA: tissue IHC). Confirm that the expected cells are present, then check retrieval, antibody incubation and detection controls; a single blank section does not establish biological absence (general IHC practice). |
💡Expected ACADL appearanceCall a result positive when High staining resolves as a cytoplasmic mitochondrial pattern in the listed thyroid, kidney or lung cell types (HPA: tissue IHC); isolated nuclear, surface or extracellular signal conflicts with ACADL's matrix location (UniProt P28330: location and topology).
| Subcellular location and topology (UniProt P28330: location and topology). | ACADL resides in the mitochondrial matrix and has no transmembrane segment (UniProt P28330); score cytoplasmic mitochondrial pattern, while treating isolated surface staining as discordant (HPA: tissue IHC; general IHC interpretation). |
| Cell specific reference pattern (HPA: tissue IHC). | Thyroid glandular cells, collecting ducts and alveolar type II cells are High; gallbladder glandular cells and hepatocytes are Medium (HPA: tissue IHC). Select the named cell compartment when assessing a tissue, since intensity differs by cell type (general IHC interpretation). |
| Evidence strength (HPA: tissue IHC reliability and antibody validation). | Tissue IHC reliability is Enhanced, while agreement between antibody staining and RNA expression is described as medium (HPA: tissue IHC). HPA011990 has Enhanced IHC validation (HPA: antibody record); retain morphology and controls when judging an individual section (general IHC practice). |
| Mature protein annotation (UniProt P28330: processing). | The annotated chain spans residues 31–430 of a 430 residue precursor (UniProt P28330). This does not identify the IHC antibody's epitope or predict retrieval performance; interpret staining from observed location and controls (general IHC practice). |
| IF evidence boundary (HPA: subcellular record and antibody record). | HPA lists no main ICC/IF location, no cell lines with ICC/IF images and no ICC status for HPA011990 (HPA: subcellular and antibody records). The tissue IHC pattern can guide an IF expectation but does not validate an IF result (HPA: tissue IHC; general IF interpretation). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known positive cells show no chromogenic signal (HPA: tissue IHC, High reference cells). | The reference compartment may be absent from the section, or an IHC workflow step may have failed (general IHC practice). | Verify the cell type on the counterstain; check a positive control, antigen retrieval record, primary antibody incubation and detection reagents (general IHC practice). |
| The section is diffusely brown and cell specific scoring is difficult (general IHC interpretation). | Nonspecific antibody binding, endogenous detection activity or excessive chromogen development may contribute (general IHC practice). | Compare a negative detection control; review blocking, wash stringency and chromogen development, then reassess the mitochondrial pattern (general IHC practice; HPA: tissue IHC profile). |
| Signal appears mainly nuclear or along cell membranes (general IHC observation). | The distribution disagrees with mitochondrial matrix ACADL and its lack of a transmembrane segment (UniProt P28330: location and topology). | Check morphology at higher magnification and compare positive and negative controls before assigning the stain to ACADL (general IHC practice). |
| Adipocytes or bone marrow hematopoietic cells stain strongly (HPA: tissue IHC, Not detected reference cells). | Cell misidentification, nonspecific staining or endogenous detection activity may explain a discordant result (general IHC interpretation). | Confirm the cell type, inspect negative controls and compare staining with a High reference cell population on a suitable section (general IHC practice; HPA: tissue IHC). |
| A Medium reference tissue stains less intensely than a High reference tissue (HPA: tissue IHC). | That difference can match HPA's cell specific categories; gallbladder glandular cells and hepatocytes are Medium (HPA: tissue IHC). | Score the named cells and their compartment separately; compare sections only after checking control performance and staining conditions (general IHC practice). |
| Can an ICC/IF image confirm the expected ACADL pattern? | HPA supplies no ICC/IF image cell line or main subcellular ICC/IF location, and lists no ICC status for HPA011990 (HPA: subcellular and antibody records). | Use mitochondrial localisation as an expectation from UniProt and tissue IHC, while recording IF staining as independently assessed rather than HPA validated (UniProt P28330: location; HPA: tissue IHC; general IF interpretation). |
Experiment evidence
IHC & IF images showing ACADL staining
A07206 · IHCImmunohistochemical analysis of ACADL staining in rat ovary formalin fixed paraffin embedded tissue section.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115361-ACADL/tissue/cerebral+cortex#imid_3125872
HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000115361-ACADL/tissue/thyroid+gland#imid_3125816
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115361-ACADL/tissue/colon#imid_3125782
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115361-ACADL/tissue/liver#imid_3125783
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACADL IHC & IF
🧪Run lung first: alveolar type II cells should stain (HPA: High in alveolar cells type II; UniProt P28330: expression in alveolar type II pneumocytes). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); assess neighboring cells outside the type II population on the lung slide for background without assuming they are ACADL-negative (HPA: lung cell-specific staining reported for alveolar cells type II).
Positive control tissue: Kidney (Collecting ducts, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ACADL; derive a cell-line control from the positive tissue's cell type (Collecting ducts) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a host-species- and clonality-matched isotype control, and ACADL-knockout tissue where available as a biological negative (standard IHC practice). Check for endogenous peroxidase activity in lung sections before chromogenic detection; if using IF, assess lung autofluorescence (standard IHC/IF practice).
⚠️Feasibility: The selected A07206 image documents ACADL staining in a formalin-fixed, paraffin-embedded rat ovary section, but reports no target-specific fixation window (A07206 tissue-IHC caption). Antigen retrieval requirements and any fixation effect are unreported in the supplied evidence; there is no basis here to call frozen sections or IF easier (supplied application evidence). In lung, endogenous peroxidase can complicate chromogenic interpretation, while autofluorescence can complicate IF interpretation (standard IHC/IF practice).
HPA tissue IHC evidence for ACADL
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 |
|---|---|---|---|---|
| Kidney | Collecting ducts | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACADL IHC Tips
Troubleshoot ACADL staining in paraffin sections by checking retrieval, cell identity, mitochondrial pattern, controls, and scoring before interpreting chromogenic signal.
How should I retrieve ACADL in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in retrieval buffer, then compare staining against a matched section processed in the same run (standard IHC practice). ACADL is a mitochondrial matrix protein whose annotated mature chain spans residues 31–430, so assess cytoplasmic mitochondrial-pattern signal rather than nuclear staining (UniProt P28330). If signal remains weak, vary retrieval duration on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Judge the result by cell-specific staining and background together, since stronger overall color alone does not establish improved specificity (standard IHC practice).
Could fixation explain weak ACADL staining in my paraffin sections?
ACADL-specific sensitivity to fixation has not been established by the supplied evidence, so treat fixation as an experimental variable rather than a known cause of weak signal (supplied evidence). The A07206 image caption documents staining in formalin-fixed, paraffin-embedded rat ovary but gives no comparison across fixation conditions (A07206 image caption). Record fixative, fixation interval, processing schedule, and section age for each block before comparing staining intensity (standard IHC practice). Run adjacent sections from well-documented blocks together using the same retrieval and detection conditions to isolate processing differences (standard IHC practice). Check morphology and internal staining patterns before attributing a negative section to altered ACADL abundance (standard IHC practice).
What ACADL staining pattern should I expect in tissue sections?
Expect cytoplasmic staining with a mitochondrial pattern, consistent with ACADL's mitochondrial matrix location and lack of an annotated transmembrane segment (HPA tissue IHC; UniProt P28330). In lung, examine alveolar type II cells because ACADL protein expression is reported there and HPA rates their staining high (UniProt P28330; HPA: High in alveolar cells type II). Collecting ducts in kidney and glandular cells in thyroid are additional high-staining reference populations (HPA tissue IHC). Compare the signal with tissue architecture and a negative reagent control on adjacent sections (standard IHC practice). Diffuse nuclear or luminal color without the expected cytoplasmic distribution needs further specificity checks (UniProt P28330; standard IHC practice).
How do processing and epitope position affect ACADL staining?
Check the antibody's stated immunogen or epitope against the annotated mature ACADL chain, residues 31–430, before interpreting a negative section (UniProt P28330). The record lists 0 isoforms and a processed chain beginning at residue 31; it does not establish where this antibody binds (UniProt P28330; supplied evidence). Reported modifications include acetylation and phosphorylation, so an epitope overlapping a modified residue could require separate validation rather than an assumed staining correction (UniProt P28330; standard antibody-validation practice). Compare staining with an independently validated antibody recognizing a different region if epitope-dependent detection is suspected (standard IHC practice). Keep retrieval and chromogenic detection matched when making that comparison (standard IHC practice).
How can IF help assess whether ACADL staining is cell specific?
On the separate IF/ICC guide workflow, multiplex ACADL with a validated alveolar type II cell marker when assessing lung, because that cell population expresses ACADL (UniProt P28330; standard IF practice). Choose a fluorophore and imaging channel with low tissue autofluorescence, and acquire unstained and single-label controls before interpreting overlap (standard IF practice). ACADL is in the mitochondrial matrix and has no annotated transmembrane segment, so permeabilisation must allow antibody access across the plasma and mitochondrial membranes (UniProt P28330; standard IF practice). Confirm the permeabilisation conditions with a validated mitochondrial marker while checking that cell morphology remains intact (standard IF practice). Fluorescent overlap supports localisation but does not alone establish antibody specificity (standard IF practice).
How do I reduce diffuse ACADL chromogenic background?
First inspect a no-primary control and review whether color follows tissue edges, folds, or damaged areas rather than intact cell cytoplasm (standard IHC practice). Use an appropriate protein block, thorough washes, and a secondary detection reagent suited to the specimen; adjust one variable at a time (standard IHC practice). Include a peroxidase block before DAB development and shorten development if color spreads beyond cellular boundaries (standard chromogenic IHC practice). Preserve the expected cytoplasmic mitochondrial pattern as the criterion for useful signal, especially in HPA high-staining thyroid glandular cells (HPA tissue IHC; standard IHC practice). A cleaner section with weaker total color can be easier to interpret than intense diffuse staining (standard IHC practice).
How should I score ACADL staining across tissue groups? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; for lung, alveolar type II cells are a biologically relevant population (UniProt P28330; HPA tissue IHC). Record the percentage of positive target cells and cytoplasmic intensity on a prespecified 0–3 scale, then calculate an H-score from 0–300 if intensity weighting is needed (standard IHC scoring practice). Alternatively, measure positive-cell density per mm² when cell abundance itself is the outcome (standard image-analysis practice). Normalize each readout to evaluable target-cell count or annotated target-cell area, and apply the same threshold and imaging settings across groups (standard IHC quantification practice). Exclude folds, necrosis, and section edges using criteria set before scoring (standard IHC practice).
How can I distinguish true ACADL signal from artefact?
A credible positive has cytoplasmic mitochondrial-pattern staining in an appropriate cell population, such as lung alveolar type II cells or thyroid glandular cells (HPA tissue IHC; UniProt P28330). Nuclear-only color conflicts with ACADL's annotated mitochondrial matrix location and warrants antibody or detection controls (UniProt P28330; standard IHC practice). Edge staining, necrotic debris, and color persisting in a no-primary control point toward processing or detection artefact (standard IHC practice). Check the peroxidase block when DAB appears in unexpected structures, because endogenous enzyme activity can produce chromogenic signal (standard chromogenic IHC practice). Interpret absent staining cautiously: HPA reports only medium consistency between antibody staining and RNA expression data (HPA tissue IHC).
Boster reagents
Best ACADL / Long-chain specific acyl-CoA dehydrogenase, mitochondrial IHC Antibodies
The IHC-validated anti-ACADL antibody has a staining image from rat ovary formalin-fixed paraffin-embedded tissue (A07206 image caption); listed reactivity covers human, mouse and rat (A07206 catalog).
Real IHC data
Anti-ACADL Antibody
Cat # A07206
A07206 has an IHC image from rat ovary formalin-fixed paraffin-embedded tissue (A07206 image caption). It lists IHC among its applications and human, mouse and rat reactivity (A07206 catalog).
Which to pick: For tissue IHC, choose A07206: it is a rabbit antibody listed for IHC (A07206 catalog), with staining shown in rat ovary formalin-fixed paraffin-embedded tissue (A07206 image caption). For IF/ICC, no SKU in this catalog lists that application or supplies an IF image (A07206 catalog). For work across species, A07206 lists human, mouse and rat reactivity, while its supplied IHC image shows rat tissue only (A07206 catalog; A07206 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P28330 (ACADL_HUMAN, Long-chain specific acyl-CoA dehydrogenase, mitochondrial).
- Human Protein Atlas. ACADL tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACADL subcellular location (ICC-IF): Highest expression in HeLa: 3.6 nTPM.
- Human Protein Atlas. ACADL antibody validation summary (1 antibodies).
- Prognostic stratification of chordoid meningiomas: the role of chromosome 1p/22q Loss. Acta neuropathologica communications 2026 — PMC13081255.
- ACADL plays a tumor-suppressor role by targeting Hippo/YAP signaling in hepatocellular carcinoma. NPJ precision oncology 2020 — PMC7096519.
- Atypical meningiomas with an immunohistochemical profile consistent with hypermetabolic or proliferative molecular groups show high mitotic index, chromosomal instability, and higher recurrence risk. Virchows Archiv : an international journal of pathology 2023 — PMC10326084.
- Refining prognostic stratification of atypical meningiomas: significance of chromosome 1p deletion and brain invasion. Acta neuropathologica communications 2025 — PMC11883970.
- PubMed PMID:1774065 — UniProt-cited evidence.
- PubMed PMID:1559716 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACADL (Long-chain specific acyl-CoA dehydrogenase, mitochondrial). Expected localisation Cytoplasmic staining with a mitochondrial pattern (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07206 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Collecting ducts); negative Adipose tissue. IHC-P workflow on FFPE sections. Protocol steps: 1. Sample: FFPE rat ovary formalin fixed paraffin embedded tissue (datasheet A07206); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A07206); 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 citrate buffer, pH 6.0 , 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-ACADL, 1:50-1:200 (datasheet A07206); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Cell mix affects intensity: type II alveolar cells stain high (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans 31–430 (UniProt); Regulation: Tissue and cell type dependent abundance (HPA tissue IHC). Evidence: UniProt P28330, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in HeLa: 3.6 nTPM, Datasheet A07206, PMC13081255, PMC7096519. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACADL/immunohistochemistry-ihc-guide.