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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 using the cytoplasmic mitochondrial pattern and thyroid glandular cells as a positive reference (HPA tissue IHC). Start with the catalog antibody’s 1:50–1:200 IHC dilution range (datasheet: A07206) and keep fixation consistent across sections (standard IHC practice).
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 mitochondrial pattern in tissue (HPA tissue IHC) | |
| Staining pattern | Thyroid glandular cells show a cytoplasmic mitochondrial pattern (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 | Staining and RNA expression have medium consistency (HPA tissue IHC) | |
| Regulation | Tissue-dependent; most abundant in thyroid (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms; mature chain lacks residues 1–30 (UniProt) |
Section 1
Recommended ACADL IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by two published ACADL IHC examples (PMC13081255; 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 retrieval at 95–98 °C for 20 min (page antigen retrieval); adjust after checking staining and controls.
Section 2
What Is the Expected ACADL Staining Pattern?
ACADL is a mitochondrial matrix protein with no transmembrane segment (UniProt P28330 localization and topology). In paraffin-section IHC, expect cytoplasmic staining with a mitochondrial pattern, especially in thyroid glandular cells, lung alveolar type II cells, and kidney collecting ducts (HPA tissue IHC). HPA rates the tissue staining evidence Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Discrete, granular cytoplasmic staining in thyroid glandular cells, alveolar type II cells, or collecting ducts. | This fits the reported mitochondrial pattern and High staining in those cell populations (HPA tissue IHC). Score the identified cells and their cytoplasmic pattern, since intensity alone does not establish specificity (standard IHC practice). |
| Predominantly nuclear, cell-surface, or extracellular staining with little granular cytoplasmic signal. | Those compartments do not fit a mitochondrial matrix protein with no transmembrane segment (UniProt P28330 localization and topology). Check morphology, counterstain, and detection controls before interpreting such staining as ACADL (standard IHC practice). |
| Strong staining in adipocytes or esophageal squamous epithelial cells while expected positive cells are faint. | HPA reports ACADL as Not detected in those two cell populations (HPA tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity; either can produce misleading chromogenic signal (standard IHC practice). |
| Uniform color across cells, stroma, and empty spaces, obscuring cellular detail. | A widespread deposit does not resemble HPA's cell-associated mitochondrial pattern (HPA tissue IHC). Assess background from primary antibody, detection reagents, and blocking with appropriate controls (standard IHC practice). |
| No convincing signal in thyroid glandular cells, alveolar type II cells, or collecting ducts. | These are reported High staining populations, so an all-negative run warrants investigation (HPA tissue IHC). Review tissue identity, section quality, retrieval, detection, and the IHC-validated antibody's instructions (standard IHC practice). |
💡Expected ACADL appearanceA convincing positive is granular cytoplasmic, mitochondrial-pattern staining in the reported High cell populations; dominant nuclear, surface, or uniform background color is suspect (HPA tissue IHC; UniProt P28330 localization and topology; standard IHC practice).
| Subcellular location | ACADL resides in the mitochondrial matrix and has no transmembrane segment (UniProt P28330 localization and topology). Interpret cytoplasmic mitochondrial-pattern staining in identified cells; membrane outlines are inconsistent with that annotation (HPA tissue IHC; UniProt P28330 topology). |
| Cell-type distribution | HPA reports High staining in thyroid glandular cells, alveolar type II cells, and kidney collecting ducts, but Medium staining in hepatocytes (HPA tissue IHC). Compare the same cell types across sections rather than whole-tissue color (standard IHC practice). |
| Protein processing | UniProt annotates a mature ACADL chain spanning residues 31–430 (UniProt P28330 processing). The payload gives no antibody epitope, so it cannot establish whether processing changes this assay's staining (UniProt P28330 processing; supplied antibody data). |
| Evidence and antibody scope | HPA lists IHC validation as Enhanced for HPA011990, yet describes only medium consistency with RNA expression (HPA antibody validation; HPA tissue IHC). Treat the listed cell patterns as reference observations, and assess unexpected staining with controls (standard IHC practice). |
| Endogenous chromogen-generating activity | Endogenous activity can add color independently of primary-antibody binding, depending on the detection chemistry (standard IHC practice). A primary-omission and detection-control comparison can help separate that signal from the reported cell-associated pattern (standard IHC practice; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive thyroid, lung, or kidney cells show no stain. | A failed assay step or unsuitable section is possible; these cells are reported High by HPA (HPA tissue IHC; standard IHC practice). | Verify cell identity and section quality, then check retrieval, antibody application, and detection against the catalog antibody's IHC-P instructions (standard IHC practice). |
| Signal is weak but limited to expected cells. | HPA's High categories describe reported observations, not a guaranteed intensity in every run (HPA tissue IHC; standard IHC practice). | Compare a known-positive control in the same run and review detection and counterstain before assigning an absence score (standard IHC practice). |
| Nuclei or cell borders stain more strongly than cytoplasm. | That distribution conflicts with ACADL's mitochondrial matrix location and HPA's cytoplasmic mitochondrial pattern (UniProt P28330 localization; HPA tissue IHC). | Check cellular morphology, primary-omission controls, and detection controls; do not score the misplaced signal as specific without corroboration (standard IHC practice). |
| Adipocytes or esophageal squamous epithelial cells stain strongly. | HPA records ACADL as Not detected in those populations; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; standard IHC practice). | Compare with known-positive cells and primary-omission controls, then reassess antibody specificity and detection background (standard IHC practice). |
| Chromogen covers tissue and surrounding background. | Diffuse deposit can arise from assay background and obscures the expected cell-associated pattern (standard IHC practice; HPA tissue IHC). | Inspect primary-omission and detection controls, blocking, washing, and chromogen development before scoring cells (standard IHC practice). |
| Can an IF/ICC image confirm the IHC pattern? | HPA supplies no ACADL ICC-IF image or main subcellular location in this payload (HPA subcellular record). | Use UniProt's mitochondrial matrix annotation as the localization expectation; consult the separate IF/ICC guide for that application's design (UniProt P28330 localization; HPA subcellular record). |
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 strongly (HPA: High in lung alveolar cells type II). Use appendix glandular cells as the tissue negative (HPA: Not detected in appendix glandular cells); on the lung slide, neighboring cells should show background-level staining to serve as an internal contrast, but their ACADL-negative status is unverified (HPA: lung row specifies alveolar cells type II only).
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 concentration-matched isotype control appropriate to the primary antibody’s host and class, and ACADL-knockout tissue or a validated peptide-block control (standard IHC practice). For chromogenic lung IHC, block endogenous peroxidase and check pigment for signal that could be mistaken for staining (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or retrieval dependency is reported in the supplied evidence; optimize antigen retrieval for the catalog antibody rather than inferring it from tissue staining (HPA: tissue staining rows; datasheet conditions not supplied). The selected A07206 caption explicitly describes a formalin-fixed, paraffin-embedded rat ovary section, but gives no fixation duration or retrieval conditions (A07206 tissue-IHC caption). The evidence does not establish that frozen sections or IF are easier for ACADL; in lung, distinguish chromogen from endogenous pigment when scoring alveolar cells (standard IHC 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 chromogenic IHC in paraffin sections by checking retrieval, cell type, mitochondrial pattern, and staining controls before interpreting signal.
How should I retrieve ACADL in paraffin sections when staining is weak?
Start ACADL paraffin IHC with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Let sections cool in retrieval buffer, then compare the same antibody dilution and detection conditions across runs (standard IHC practice). Include a positive control such as lung alveolar type II cells or kidney collecting ducts, both reported with high staining (HPA tissue IHC). If signal remains weak, test a longer exposure to the same citrate buffer on paired sections, while watching for tissue damage (standard IHC practice). Judge improvement by the expected cytoplasmic mitochondrial pattern, rather than total brown staining alone (HPA tissue IHC; UniProt P28330 localisation).
How can I troubleshoot variable ACADL staining across differently fixed paraffin blocks?
ACADL-specific sensitivity to fixation conditions is unknown because no fixation comparison is supplied (evidence payload). For each block, record fixative, fixation duration, processing history, and section age before comparing staining intensity (standard IHC practice). Keep retrieval at citrate pH 6.0, 95–98 °C for 20 min while assessing block-to-block variation (page retrieval rule). Stain paired sections in one run with a shared positive control to distinguish processing variation from detection variation (standard IHC practice). Interpret any difference cautiously: neither the reported mitochondrial location nor the annotated modified residues establish a particular fixation effect (UniProt P28330 localisation and modified residues).
What ACADL staining pattern should I expect in chromogenic tissue IHC?
Expect cytoplasmic staining with a mitochondrial pattern, consistent with ACADL residing in the mitochondrial matrix (HPA tissue IHC; UniProt P28330 localisation). Examine cell bodies at high magnification and compare fine granular staining with the diffuse counterstain and any coarse pigment (standard IHC practice). Lung alveolar type II cells, kidney collecting ducts, and thyroid glandular cells are reported as high-staining populations (HPA tissue IHC). Score the fraction and intensity of appropriately localised cells separately from stromal or luminal deposits (standard IHC practice). If staining appears predominantly nuclear or outlines the plasma membrane, recheck controls and detection conditions before assigning it to ACADL (UniProt P28330 localisation; standard IHC practice).
Could ACADL processing or epitope choice explain inconsistent tissue staining?
The record lists 0 isoforms and a processed chain spanning residues 31–430, so an isoform-specific staining explanation is unsupported here (UniProt P28330 processing and isoforms). The protein has no annotated transmembrane segment and resides in the mitochondrial matrix (UniProt P28330 topology and localisation). Check whether the antibody's documented immunogen overlaps the processed chain before interpreting a negative section; that immunogen is not supplied here (UniProt P28330 processing; evidence payload). Several modified residues are annotated, including positions 42 and 54, but their effects on this antibody's binding are unknown (UniProt P28330 modified residues). Compare independent positive tissue controls under identical retrieval conditions before attributing discrepancies to epitope accessibility (standard IHC practice).
How should I assess ACADL by multiplex IF alongside this IHC guide?
For the separate IF/ICC workflow, pair ACADL with a validated marker of lung alveolar type II cells when testing that reported positive population (HPA tissue IHC; UniProt P28330 tissue specificity). Because tissue autofluorescence can confound dim signal, inspect unstained sections and assign ACADL a fluorophore in a channel with lower background (standard IF practice). ACADL is in the mitochondrial matrix and has no transmembrane segment, so permeabilisation must allow antibody access across cellular and mitochondrial membranes (UniProt P28330 localisation and topology). Optimise permeabilisation on paired specimens and confirm that the cell marker and ACADL remain distinguishable (standard IF practice). The supplied HPA subcellular payload lists 0 ICC/IF image cell lines, so it does not establish an IF staining pattern for this antibody (HPA subcellular).
How do I reduce nonspecific brown staining in ACADL IHC?
First compare a no-primary control with the stained section to locate signal introduced by the detection chemistry (standard IHC practice). Quench endogenous peroxidase before an HRP and DAB workflow, and use an appropriate protein block for nonspecific antibody binding (standard chromogenic IHC practice). If background rises after citrate pH 6.0 retrieval for 20 min, compare paired sections while keeping detection exposure constant (page retrieval rule; standard IHC practice). Titrate the IHC-validated antibody only within its documented range, which is not provided in this payload (evidence payload). Retain staining only when it resolves into the reported cytoplasmic mitochondrial pattern in plausible cells and exceeds no-primary background (HPA tissue IHC; standard IHC practice).
How should I quantify ACADL staining across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before scoring, since ACADL staining differs among cell types within organs (HPA tissue IHC). For chromogenic IHC, report an H-score from intensity categories 0–3 and the percentage of cells in each category, or report percentage positive cells alone (standard IHC practice). Normalize counts to the number of evaluable target cells; if measuring positive-cell density, use cells per mm² of viable tissue (standard IHC practice). Exclude folds, necrosis, section edges, and blank spaces using the same rules for every sample (standard IHC practice). Keep retrieval, imaging, thresholds, and the positive control consistent, and score cytoplasmic signal separately from off-pattern deposits (page retrieval rule; HPA tissue IHC; standard IHC practice).
When is ACADL staining convincing rather than an IHC artefact?
A convincing result shows cytoplasmic mitochondrial-pattern staining in a plausible cell population, such as lung alveolar type II cells or kidney collecting ducts (HPA tissue IHC; UniProt P28330 localisation). Check that the pattern repeats away from section edges and damaged or necrotic areas, and that the no-primary control stays clear (standard IHC practice). Predominantly nuclear or surface staining conflicts with the annotated mitochondrial matrix location and warrants further validation (UniProt P28330 localisation). Diffuse brown signal shared by control sections can reflect endogenous peroxidase or detection background in an HRP and DAB workflow (standard chromogenic IHC practice). Interpret a negative result against the same-run positive control, because HPA reports some cell populations as not detected and rates tissue staining reliability as Enhanced with medium RNA agreement (HPA tissue IHC).
Boster reagents
Best ACADL / Long-chain specific acyl-CoA dehydrogenase, mitochondrial IHC Antibodies
The IHC-listed anti-ACADL antibody A07206 has a rat ovary staining image from a formalin-fixed, paraffin-embedded section (catalog image caption) and listed human, mouse, and rat reactivity (catalog).
Real IHC data
Anti-ACADL Antibody
Cat # A07206
A07206 will render with its IHC image of a formalin-fixed, paraffin-embedded rat ovary section (catalog image caption). Its listed application is IHC, with human, mouse, and rat reactivity (catalog applications and reactivity).
Which to pick: For tissue IHC, choose A07206: its own image documents rat ovary staining in a formalin-fixed, paraffin-embedded section (catalog image caption), and the listed IHC dilution is 1:50–1:200 (datasheet). For cross-species work, A07206 lists human, mouse, and rat reactivity, while its supplied IHC image shows rat tissue only (catalog reactivity; catalog image caption). For IF/ICC, the payload lists no IF application or IF image for A07206 (catalog applications and image data); its host is rabbit and clonality is unreported (catalog).
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 mitochondrial pattern in tissue (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: Staining and RNA expression have medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain lacks residues 1–30 (UniProt); Regulation: Tissue-dependent; most abundant in thyroid (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.