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ACAD11 / Acyl-CoA dehydrogenase family member 11 · IHC design guide
Design Immunohistochemistry for ACAD11
Plan chromogenic ACAD11 IHC in paraffin sections around the granular cytoplasmic staining reported in brain, kidney and liver (HPA tissue IHC). This guide covers fixation consistency, the catalog antibody’s 1:100–1:300 IHC dilution (datasheet), and interpretation of cell-specific staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACAD11 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 | Tissue: granular cytoplasm (HPA tissue IHC); annotated: peroxisome and mitochondrial membrane (UniProt) | |
| Staining pattern | Granular cytoplasm in hepatocytes and renal tubules (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Cerebellar staining favors granular-layer processes (HPA tissue IHC) | |
| Regulation | Tissue-dependent staining intensity (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; antibody epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended ACAD11 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by a published ACAD11 IHC protocol for human cerebellum and muscle (PMC3073726).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10875-1) |
| Fixation | Image fixative and duration unreported (datasheet A10875-1); 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-ACAD11, 1:100-1:300 (datasheet A10875-1) |
| 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 | ACAD11-positive staining in enterocytes - Microvilli of appendix (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in selected tissues, most abundant in brain, kidney and liver. 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); the article does not specify retrieval (PMC3073726).
Section 2
What Is the Expected ACAD11 Staining Pattern?
ACAD11 is reported at peroxisomes and, in some settings, at the mitochondrial membrane; it has no transmembrane segment (UniProt Q709F0). In paraffin-section IHC, expect granular cytoplasmic staining, especially in cerebellar granular-layer processes, kidney proximal tubule cells and hepatocytes (HPA tissue IHC). HPA rates its IHC evidence Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC).
| Granular cytoplasmic staining is strong in cerebellar granular-layer processes and moderate in kidney proximal tubule cell bodies or hepatocytes. | This matches the reported cell-level pattern: High in cerebellar processes and Medium in those kidney and liver cells (HPA tissue IHC). Score each cell population and its compartment; intensity need not match across tissues. |
| Signal is predominantly nuclear or forms a continuous cell-surface outline instead of cytoplasmic granules. | That distribution differs from the reported granular cytoplasmic IHC pattern (HPA tissue IHC). UniProt reports no transmembrane segment, although membrane association occurs in some settings (UniProt Q709F0). Check specificity before calling it ACAD11. |
| Strong staining appears in adipocytes or bronchial respiratory epithelial cells. | HPA reports ACAD11 as Not detected in those cell populations (HPA tissue IHC). Consider nonspecific antibody binding or endogenous detection activity; compare a matched control. An HPA negative is a reference observation, not proof that every specimen must be negative. |
| Brown signal spreads across cells or tissue without a discernible granular cytoplasmic pattern. | Diffuse signal cannot be confidently assigned to ACAD11's reported pattern (HPA tissue IHC). As a general IHC check, inspect the no-primary control and assess blocking, antibody concentration and chromogen development before scoring. |
| A cerebellar granular layer, kidney proximal tubule region or hepatocyte region has no detectable signal. | Each is a reported positive cell population, although kidney and liver are rated Medium rather than High (HPA tissue IHC). Confirm that the intended cells are present, then check the staining run and a positive control before interpreting absence as biological. |
💡Expected ACAD11 appearanceCall the IHC result consistent with ACAD11 when discrete granular cytoplasmic signal is High in cerebellar granular-layer processes or Medium in kidney proximal tubule cell bodies and hepatocytes; predominantly nuclear, continuous surface or indiscriminate diffuse staining warrants investigation (HPA tissue IHC; UniProt Q709F0 topology).
| Which tissue gives the clearest reference pattern? | Cerebellar granular-layer processes are rated High; proximal tubule cell bodies and hepatocytes are rated Medium (HPA tissue IHC). Use the named cell population when judging a section, rather than its tissue-wide color. |
| How far can organelle identity be inferred? | UniProt lists peroxisomes and mitochondrial membrane association, with observations differing by tissue and cell context (UniProt Q709F0). Granular cytoplasmic chromogen alone does not distinguish those organelles; organelle assignment needs separate evidence. |
| How strong is the IHC pattern evidence? | HPA reports Enhanced IHC validation for HPA048317 and describes medium consistency between antibody staining and RNA expression (HPA antibodies; HPA tissue IHC). Treat the mapped pattern as a reference while retaining the stated consistency limit. |
| Do isoforms or processing define a different staining pattern? | UniProt lists 3 isoforms and one annotated chain spanning residues 1–780 (UniProt Q709F0). The supplied evidence does not establish which isoforms the antibody detects, so staining cannot identify an isoform. |
| Could chromogenic detection create a misleading positive? | Endogenous enzyme activity can contribute color in routine chromogenic IHC (general IHC practice). A matched no-primary control helps identify detection-derived signal; this is a general workflow check, not an ACAD11-specific HPA finding. |
| IF/ICC Q&A: is a cultured-cell pattern established? | No HPA ICC-IF images or main subcellular location are available for ACAD11, and the listed antibody has no ICC validation entry (HPA subcellular; HPA antibodies). The tissue IHC pattern therefore does not establish an expected IF/ICC image. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal appears in a section containing the reported positive cells. | A staining-run problem is possible; kidney and liver positives are only Medium on the HPA scale (HPA tissue IHC). | Verify the relevant cells on the section and inspect a concurrent positive control. As general IHC practice, review retrieval, primary incubation and detection if that control also fails; no ACAD11-specific retrieval effect is established. |
| The positive control stains, but the test section lacks visible granules. | The sampled cells may differ from the HPA-positive populations, or the test specimen may genuinely differ (HPA tissue IHC). | Confirm cell identity and morphology before calling a negative. Compare like cell populations and report the control result; avoid treating whole-tissue absence as proof of absent ACAD11. |
| Nuclei or uninterrupted cell borders dominate the signal. | This conflicts with HPA's granular cytoplasmic pattern; UniProt reports no transmembrane segment (HPA tissue IHC; UniProt Q709F0). | Check the no-primary control and review antibody concentration and detection as general IHC practice. Score ACAD11 only where the cell-level pattern is supported. |
| Unexpected strong color appears in adipocytes or bronchial respiratory epithelium. | Those cells are listed as Not detected by HPA; nonspecific binding or detection background is possible (HPA tissue IHC; general IHC practice). | Compare matched controls and examine whether color follows the expected granular cytoplasmic distribution. Record the discrepancy rather than reclassifying those cells as established positives. |
| Diffuse brown background obscures tissue boundaries. | Excess antibody, incomplete blocking or prolonged chromogen development can raise background in routine IHC (general IHC practice). | Inspect the no-primary control and adjust one general staining variable at a time. Reassess whether cytoplasmic granules remain distinct in a reported positive cell population (HPA tissue IHC). |
| Kidney or liver looks weaker than cerebellum. | That ordering is compatible with HPA's High rating for cerebellar processes and Medium ratings for proximal tubule cell bodies and hepatocytes (HPA tissue IHC). | Score the specified cells and compartment within each tissue. Investigate a run problem only if the expected pattern or a concurrent positive control is also lost. |
Experiment evidence
IHC & IF images showing ACAD11 staining
A10875-1 · IHCImmunohistochemistryt analysis of paraffin-embedded human breast carcinoma, using ACAD11 Antibody. The lane on the right is blocked with the
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000240303-ACAD11/tissue/cerebral+cortex#imid_14074114
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000240303-ACAD11/tissue/cerebellum#imid_14074117
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000240303-ACAD11/tissue/colon#imid_14052832
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000240303-ACAD11/tissue/liver#imid_14052825
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACAD11 IHC & IF
🧪Run appendix first and look for staining in enterocyte microvilli (HPA: High in appendix enterocyte microvilli). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the appendix slide, nuclei and nonmicrovillar compartments should lack the same apical enrichment, without assuming that every other cell is ACAD11-negative.
Positive control tissue: Appendix (Enterocytes - Microvilli, 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 ACAD11; derive a cell-line control from the positive tissue's cell type (Enterocytes - Microvilli) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality (standard IHC practice). Use ACAD11 knockout material if available, or a peptide-block control as shown for the catalog antibody (A10875-1 caption: ACAD11 peptide block); for chromogenic appendix IHC, quench endogenous peroxidase and check background in the no-primary control (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10875-1 paraffin-section caption does not state a fixative (A10875-1 caption: fixative not stated). ACAD11-specific antigen-retrieval dependence is unreported; optimize retrieval with the catalog antibody’s IHC-P protocol, and do not assume frozen sections or IF are easier (standard IHC practice). In appendix, preserve and assess the microvillar border when scoring because the expected positive compartment is narrow and apical (HPA: High in appendix enterocyte microvilli).
HPA tissue IHC evidence for ACAD11
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 |
|---|---|---|---|---|
| Appendix | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Cerebellum | Processes in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACAD11 IHC Tips
Troubleshoot ACAD11 staining in paraffin sections by checking retrieval, cellular pattern, controls, and scoring before interpreting chromogenic signal.
What retrieval conditions should I start with for weak ACAD11 staining?
Use citrate buffer at pH 6.0 for heat-induced antigen retrieval at 95–98 °C for 20 min in ACAD11 paraffin-section IHC (page retrieval rule). Cool sections consistently, then compare a retrieval-treated section with an untreated section using the same antibody incubation and chromogenic development (standard IHC practice). If staining remains weak, test a second retrieval condition as a fallback on adjacent sections, while checking whether tissue morphology or background deteriorates (standard IHC practice). Judge improvement in the expected granular cytoplasmic pattern in brain, kidney, or liver, rather than by overall stain intensity alone (HPA: tissue IHC profile).
Could fixation explain weak or uneven ACAD11 staining?
ACAD11-specific sensitivity to fixation is unknown from the supplied evidence; the catalog image identifies paraffin-embedded breast carcinoma but does not state its fixative (catalog caption: A10875-1). Record the fixative and fixation duration for each specimen, and compare sections processed together before attributing a difference to ACAD11 expression (standard IHC practice). Keep section thickness, citrate retrieval at pH 6.0, and chromogenic development constant during that comparison (page retrieval rule; standard IHC practice). Inspect tissue preservation and staining near section edges, since uneven processing can make a weak or patchy result difficult to interpret (standard IHC practice).
Which ACAD11 staining pattern fits the known biology?
Look for granular cytoplasmic staining in appropriate cells, since that is the reported tissue pattern (HPA: tissue IHC profile). ACAD11 is associated with peroxisomes and has been detected at mitochondrial membranes in some sampled contexts, while no transmembrane segment is annotated (UniProt Q709F0: subcellular location and topology). In kidney, examine proximal tubule cell bodies; in liver, examine hepatocytes, and assess the cellular pattern rather than treating diffuse brown pigment as equivalent staining (HPA: kidney and liver IHC; standard IHC practice). Do not require every specimen to show an identical organelle pattern, because reported mitochondrial association varies by sampled context (UniProt Q709F0: subcellular location).
How should I assess a discrepant ACAD11 epitope result?
Check which sequence the antibody recognizes before comparing stained specimens: ACAD11 has 3 annotated isoforms, and the supplied caption does not identify the catalog antibody’s epitope (UniProt Q709F0: isoforms; catalog caption: A10875-1). An epitope absent from an isoform could change staining without a corresponding change in total ACAD11, so verify the sequence against each isoform before making that inference (UniProt Q709F0: isoforms; standard antibody interpretation). Lys 177, Tyr 324, and Lys 391 have annotated modifications, but their effect on this antibody’s binding is unknown (UniProt Q709F0: modified residues). Compare staining with an independently mapped epitope or an orthogonal expression measure when the distinction matters (standard IHC practice).
How can I investigate ACAD11 localisation by IF alongside tissue IHC?
Treat IF as a separate assay: the supplied HPA subcellular record lists no ICC/IF images, so the tissue IHC pattern does not establish an IF staining result (HPA: subcellular record; HPA: tissue IHC profile). Multiplex ACAD11 with a marker identifying the cell population under examination, such as a proximal tubule marker in kidney, and first confirm that each channel stains as expected alone (HPA: kidney IHC; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence in each channel, and include single-label controls for bleed-through (standard IF practice). Set permeabilisation according to the mapped epitope’s accessible side and verify that treatment preserves the relevant cellular structures (standard IF practice).
How do I reduce diffuse brown background without losing ACAD11 signal?
First compare the stained section with a no-primary control and inspect whether the brown product follows cells or pools in damaged areas (standard chromogenic IHC practice). Block endogenous peroxidase before detection, for example with 3% hydrogen peroxide for 10 min, and check the detection system’s instructions for tissue compatibility (standard chromogenic IHC practice). Optimise the primary antibody concentration and chromogen development on adjacent sections rather than assuming stronger colour means more ACAD11 (standard IHC practice). Retain a section with the expected granular cytoplasmic pattern as a reference; the selected catalog image also reports peptide blocking, though its fixative is unstated (HPA: tissue IHC profile; catalog caption: A10875-1).
What should I score when comparing ACAD11 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring, since ACAD11 staining varies by tissue and cell type (HPA: tissue IHC profile). For a selected population, record the percentage of positive cells and an intensity-based H-score on a consistent 0–300 scale, or measure positive area per mm² when cell boundaries cannot be scored reliably (standard IHC scoring practice). Normalise to the number of evaluable cells or viable tissue area, and apply the same background threshold and development conditions across specimens (standard IHC practice). Report cell-specific results separately, such as proximal tubule cell bodies versus other kidney structures, rather than a whole-section average (HPA: kidney IHC; standard IHC practice).
When should an apparent ACAD11-positive area be considered artefactual?
Give greater weight to cellular, granular cytoplasmic signal in an expected population than to staining confined to a cut edge, necrotic area, or tissue debris (HPA: tissue IHC profile; standard IHC practice). In kidney, proximal tubule cell bodies are a reported staining site; an unexpected structure needs its own controls before being called ACAD11-positive (HPA: kidney IHC). Compare serial sections with a no-primary control to identify nonspecific detection or residual endogenous enzyme activity, and check whether signal persists away from damaged regions (standard chromogenic IHC practice). Interpret discrepancies cautiously because HPA reports only medium consistency between antibody staining and RNA expression data despite its Enhanced reliability designation (HPA: reliability description).
Boster reagents
Best ACAD11 / Acyl-CoA dehydrogenase family member 11 IHC Antibodies
A10875-1 has an IHC image of paraffin-embedded human breast carcinoma (IHC image caption). IF is listed without an IF image (catalog: applications; IF image alts). Reactivity includes Human, Mouse, and Rat (catalog: reactivity).
Real IHC data
Anti-ACAD-11 Antibody
Cat # A10875-1
A10875-1 has an IHC image of paraffin-embedded human breast carcinoma with a peptide-blocked comparator (IHC image caption). It lists IHC and IF applications and Human, Mouse, and Rat reactivity; no IF image is supplied (catalog: applications, reactivity, IF image alts).
Which to pick: Choose A10875-1 for tissue IHC because its image shows staining in a paraffin-embedded human breast carcinoma section; the fixative is unreported (IHC image caption). For IF, A10875-1 lists that application, but no IF image or ICC application is supplied (catalog: applications, IF image alts). It is also the listed cross-species option for Human, Mouse, and Rat, though the supplied IHC image shows only human tissue (catalog: reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q709F0 (ACD11_HUMAN, Acyl-CoA dehydrogenase family member 11).
- Human Protein Atlas. ACAD11 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACAD11 subcellular location (ICC-IF): Highest expression in HTERT-RPE1: 17.9 nTPM.
- Human Protein Atlas. ACAD11 antibody validation summary (1 antibodies).
- Identification and characterization of new long chain acyl-CoA dehydrogenases. Molecular genetics and metabolism 2011 — PMC3073726.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACAD11 (Acyl-CoA dehydrogenase family member 11). Expected localisation Tissue: granular cytoplasm (HPA tissue IHC); annotated: peroxisome and mitochondrial membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10875-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Enterocytes - Microvilli); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10875-1); 2. Fixation: Image fixative and duration unreported (datasheet A10875-1); 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-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-ACAD11, 1:100-1:300 (datasheet A10875-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Cerebellar staining favors granular-layer processes (HPA tissue IHC); Isoform / epitope: 3 isoforms; antibody epitope coverage is unspecified (UniProt; datasheet); Regulation: Tissue-dependent staining intensity (HPA tissue IHC). Evidence: UniProt Q709F0, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in HTERT-RPE1: 17.9 nTPM, Datasheet A10875-1, PMC3073726. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACAD11/immunohistochemistry-ihc-guide.