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ACSL1 / Long-chain-fatty-acid--CoA ligase 1 · IHC design guide
Design Immunohistochemistry for ACSL1
Plan ACSL1 IHC in paraffin sections around cytoplasmic staining in adipocytes, hepatocytes and kidney distal tubules (HPA tissue IHC). Start the IHC-validated antibody at 0.5–1 µg/mL (datasheet PB10025), then score cell types against appropriate controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ACSL1 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasm of adipocytes, hepatocytes and distal tubules (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB10025) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Appendix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB10025) | |
| Caveat | Adipocyte-rich regions can dominate tissue scoring (HPA tissue IHC) | |
| Regulation | Tissue-dependent expression (UniProt) | |
| Isoform / epitope | Three isoforms; cytoplasmic epitope coverage is unknown (UniProt) |
Section 1
Recommended ACSL1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet PB10025) is followed by published ACSL1 staining methods for mouse brain and kidney sections (PMC13073305; PMC6735294).
| Sample | Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet PB10025) |
| Fixation | Image fixative and duration unreported (datasheet PB10025); 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 PB10025); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB10025) |
| Primary antibody | Rabbit anti-ACSL1, 0.5-1μg/ml (datasheet PB10025) |
| Primary incubation | Overnight at 4 °C (datasheet PB10025) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB10025) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ACSL1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, mainly in adipose tissue, sebaceous glands, liver and kidney. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet PB10025). The brain study used pH 9.0 with fluorescent detection (PMC13073305).
Section 2
What Is the Expected ACSL1 Staining Pattern?
ACSL1 staining in paraffin sections should appear mainly cytoplasmic, with strong signal in adipocytes, hepatocytes and kidney distal tubules (HPA: Enhanced tissue IHC; HPA: High in these cells). ACSL1 is anchored in mitochondrial outer, peroxisomal, microsomal and endoplasmic reticulum membranes, with most of the protein facing the cytoplasm (UniProt P33121 topology). The expected slide pattern is cellular staining, rather than a sharp outline of the cell surface.
| Strong cytoplasmic staining in hepatocytes, adipocytes or kidney distal tubules. | This fits the reported high IHC signal in those specific cell populations (HPA: High). Compare stained cells with nearby tissue structures; HPA's assessment is cell specific, so a positive kidney section does not imply equally strong staining throughout every tubule. |
| Predominantly nuclear staining, or staining restricted to the cell surface. | That distribution conflicts with the reported cytoplasmic tissue pattern and membrane topology (HPA: tissue IHC profile; UniProt P33121 topology). Treat it as suspect until a positive tissue control and an independently validated antibody support the same compartment. |
| Strong staining in a cell population reported as not detected. | For example, appendix endocrine cells and caudate glial cells are reported as not detected (HPA: tissue IHC). Cross-reactivity or endogenous chromogenic activity is possible (general IHC practice). Assess the named cells, since a negative call for one population does not classify the whole tissue. |
| Diffuse color across cells and surrounding tissue, with little cell-level contrast. | This does not resemble the reported cytoplasmic staining in defined cell populations (HPA: tissue IHC profile). Nonspecific antibody binding or detection background may obscure interpretation (general IHC practice); compare a no-primary control before scoring weak signal. |
| No visible staining in hepatocytes or adipocytes. | Both are reported as high staining populations (HPA: High in liver hepatocytes and adipose adipocytes), so an absent signal calls the IHC run into question. Check tissue preservation and positive controls, then review retrieval and detection steps (general IHC practice). |
💡Expected ACSL1 appearanceCall a result positive when defined cells, especially adipocytes, hepatocytes or kidney distal tubules, show clear cytoplasmic staining at the reported high level (HPA: High); isolated nuclear color or uniform diffuse deposit is suspect against the reported tissue pattern (HPA: cytoplasmic profile).
| Subcellular distribution | ACSL1 occupies several intracellular membranes and has a cytoplasm-facing region from residues 46–698 (UniProt P33121 topology). A chromogenic IHC image may read as cytoplasmic rather than resolving each organelle (HPA: tissue IHC profile); do not infer one specific organelle from brown cytoplasmic color alone. |
| Choice of comparison cells | High IHC signal is reported in adipocytes, hepatocytes, cardiomyocytes, kidney distal tubules and lung alveolar type II cells (HPA: High). HPA also reports selected cells as low or not detected; compare the same cell type and compartment when judging an unexpected result. |
| Antibody validation | HPA rates ACSL1 tissue IHC reliability Enhanced and lists two antibodies with Enhanced IHC validation, HPA011316 and HPA011964 (HPA: tissue IHC; HPA: antibodies). That supports the reported pattern, but agreement with HPA alone does not prove that every signal from a different antibody is specific. |
| Isoforms and epitope | UniProt lists three ACSL1 isoforms (UniProt P33121). Without an epitope location for the antibody in this payload, the record cannot predict whether each isoform is detected or explain a discrepant tissue result by isoform choice. |
| IF/ICC Q&A: should vesicle signal match IHC? | HPA calls the ICC-IF location vesicles, with uncertain confidence, in images from A-431, RT-4 and U2OS (HPA: subcellular ICC-IF). This observation does not establish an organelle-specific pattern in paraffin tissue or replace the cytoplasmic IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive liver section has no hepatocyte signal. | Hepatocytes are reported as high staining (HPA: liver IHC); an absent signal may reflect an assay failure or an unsuitable specimen (general IHC practice). | Confirm that the tissue contains intact hepatocytes; check the positive control, antibody preparation, retrieval record and chromogenic detection steps before interpreting experimental negatives. |
| The slide shows nuclear color but little cytoplasmic color. | A nuclear-only pattern disagrees with ACSL1's intracellular membrane topology and HPA's cytoplasmic tissue profile (UniProt P33121 topology; HPA: tissue IHC). | Review the counterstain and no-primary control; seek the expected cytoplasmic signal in an HPA high cell population before calling nuclear color ACSL1. |
| Many cell types stain equally strongly in a mixed tissue. | Uniform staining weakens the expected cell-level contrast (HPA: tissue IHC). Nonspecific binding or endogenous detection activity can also produce color (general IHC practice). | Compare HPA high and not-detected cell populations by name; inspect no-primary and detection controls, then optimize blocking or antibody concentration if background persists (general IHC practice). |
| Diffuse brown deposit obscures cell boundaries. | Diffuse deposit prevents a confident match to the reported cytoplasmic pattern (HPA: tissue IHC profile); detection background is a possible explanation (general IHC practice). | Check a no-primary control and review blocking, washing and chromogen development (general IHC practice). Score only cell-associated signal that remains distinct from background. |
| Kidney staining is present, but distal tubules are weak. | High kidney staining is assigned specifically to distal tubules (HPA: kidney IHC). A positive section-level impression can therefore conceal a mismatch in the relevant cells. | Identify distal tubules on the section and compare their cytoplasmic signal with a positive control; review assay performance if those cells remain weak (general IHC practice). |
| An HPA not-detected comparator appears positive. | HPA's negative observations apply to named populations, such as appendix endocrine cells or caudate glial cells (HPA: tissue IHC); color elsewhere in those tissues does not contradict that call. | Verify the stained cell identity first. If the named population itself is positive, assess no-primary and independent-antibody controls before attributing the color to ACSL1 (general IHC practice). |
Experiment evidence
IHC & IF images showing ACSL1 staining
PB10025 · IHCIHC analysis of ACSL1 using anti-ACSL1 antibody (PB10025). ACSL1 was detected in a paraffin-embedded section of mouse kidney tissue. Heat me
PB10025 · IHCIHC analysis of ACSL1 using anti-ACSL1 antibody (PB10025). ACSL1 was detected in a paraffin-embedded section of rat kidney tissue. Heat medi
PB10025 · IFIF analysis of ACSL1 using anti-ACSL1antibody (PB10025). ACSL1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151726-ACSL1/tissue/cerebral+cortex#imid_3279552
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151726-ACSL1/tissue/colon#imid_3279486
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151726-ACSL1/tissue/liver#imid_3279479
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151726-ACSL1/tissue/kidney#imid_3279480
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA011317; green = ACSL1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA011317; green = ACSL1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ACSL1 IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain (HPA: High in cardiomyocytes). Use cervix glandular cells as the negative tissue (HPA: Not detected in cervix glandular cells); on the heart slide, compare non-cardiomyocyte cells for background, without assuming they lack ACSL1 (HPA: cardiomyocytes are the reported positive cell type).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ACSL1 in A-431, RT-4, U2OS, with annotated localisation: Vesicles (uncertain) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), matched rabbit IgG isotype, and ACSL1-knockout tissue controls (caption: rabbit anti-ACSL1; standard IHC practice). Check endogenous peroxidase and biotin background in heart sections when using the caption’s biotin-based DAB detection (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected PB10025 paraffin-section caption does not state a fixative (caption: fixative not stated). The reported IHC result used heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required; it also does not establish that frozen sections or IF are easier (caption: mouse kidney paraffin section with EDTA retrieval). For heart sections, blood-associated peroxidase can produce chromogenic background, so assess it with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for ACSL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACSL1 IHC Tips
Troubleshoot ACSL1 staining in paraffin sections by checking retrieval, cellular distribution and controls before interpreting DAB intensity (datasheet PB10025; HPA tissue IHC).
How should I troubleshoot weak ACSL1 staining after antigen retrieval?
Use heat-mediated EDTA retrieval at pH 8.0 for paraffin-section ACSL1 staining (datasheet PB10025). The selected kidney image used this retrieval before 1 μg/mL catalog antibody overnight at 4°C and DAB development (caption PB10025). If staining is weak, check deparaffinisation and even heating, then compare retrieval durations while holding antibody concentration constant (standard IHC practice). Evaluate signal in kidney distal tubules, a reported high-staining cell population, against nearby background (HPA: kidney distal tubules High). Record retrieval time and DAB development time for each section so that a change in signal has an identifiable cause (standard IHC practice).
Does fixation explain weak or uneven ACSL1 staining?
The selected ACSL1 kidney image identifies a paraffin section but does not report its fixative or fixation duration (caption PB10025). Consequently, target-specific fixation sensitivity is unknown; do not infer it from tissue staining patterns or protein topology (caption PB10025; HPA tissue IHC; UniProt P33121 topology). Compare sections with documented processing histories under the same EDTA pH 8.0 retrieval and antibody conditions (datasheet PB10025; standard IHC practice). Include a positive control section in each run, and inspect both control and test sections for uneven staining or damaged morphology (standard IHC practice). If processing histories differ, report that limitation alongside the staining result (standard IHC practice).
Which ACSL1 staining pattern is plausible in paraffin sections?
Expect predominantly cytoplasmic staining at tissue-image resolution, with kidney distal tubules among the reported high-staining cells (HPA tissue IHC: cytoplasmic profile; kidney distal tubules High). ACSL1 is associated with mitochondrial outer, peroxisomal, microsomal and endoplasmic reticulum membranes (UniProt P33121 subcellular location). Its transmembrane segment spans residues 25–45, while residues 46–698 are cytoplasmic (UniProt P33121 topology). Do not claim that a DAB pattern identifies one of those organelles without an independent compartment marker (standard IHC practice). Check suspicious nuclear or extracellular staining against a no-primary control and a section with preserved cell morphology (UniProt P33121 subcellular location; standard IHC practice).
Could epitope position or isoforms change my ACSL1 interpretation?
ACSL1 has 3 annotated isoforms, so establish which sequence the antibody immunogen represents before interpreting differences between samples (UniProt P33121 isoforms; standard IHC practice). The protein has a transmembrane segment at residues 25–45 and a cytoplasmic region at 46–698 (UniProt P33121 topology). Annotated modifications include phosphorylation at residue 84, glycosylation at 135 and acetylation at 207 and 356 (UniProt P33121 modifications/glycosylation). These annotations identify features to check against the mapped epitope; they do not establish an effect on this antibody’s staining (UniProt P33121; standard IHC practice). If the epitope is unavailable, report isoform recognition as unresolved and compare antibody results with an independent measure (standard IHC practice).
How can IF help assess an ambiguous ACSL1 IHC pattern?
For a separate IF assessment, pair ACSL1 with a marker identifying the expected cell type, such as a validated distal-tubule marker in kidney (HPA: kidney distal tubules High; standard IF practice). Check an unstained section before choosing fluorophores, and select channels that separate specific signal from tissue autofluorescence (standard IF practice). If the antibody epitope lies within residues 46–698, allow permeabilisation appropriate for access to that cytoplasmic region (UniProt P33121 topology; standard IF practice). Compare marker overlap with a no-primary control and consistent imaging settings across sections (standard IF practice). Treat apparent vesicles cautiously: the HPA IF location is labelled uncertain, whereas tissue IHC is described as cytoplasmic (HPA subcellular; HPA tissue IHC).
What should I check when DAB obscures ACSL1-positive cells?
First inspect a no-primary control to distinguish antibody-associated staining from background generated by the detection system (standard IHC practice). Use a peroxidase block and assess whether the biotin-based detection system contributes background before interpreting DAB intensity (standard IHC practice; caption PB10025). The selected kidney procedure used 10% goat serum blocking, a biotinylated secondary antibody and DAB (caption PB10025). If staining remains diffuse, titrate the primary around the documented 1 μg/mL condition while keeping retrieval and development time fixed (caption PB10025; standard IHC practice). Judge improvement by clearer cell boundaries in a reported positive population, such as kidney distal tubules (HPA: kidney distal tubules High; standard IHC practice).
How should I quantify ACSL1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; kidney distal tubules are a documented high-staining population (HPA: kidney distal tubules High; standard IHC practice). Record the percentage of positive cells and staining intensity, or calculate an H-score from percentages across intensity categories (standard IHC practice). If measuring stained area or cell density per mm², normalise to the viable tissue area or number of eligible cells in the same region (standard IHC practice). Keep retrieval at EDTA pH 8.0 and hold detection and imaging settings consistent across compared sections (datasheet PB10025; standard IHC practice). Exclude folds and poorly preserved areas using the same prespecified rules for every section (standard IHC practice).
How do I separate true ACSL1 staining from artefact?
Look for reproducible cytoplasmic staining in a documented positive cell population, such as kidney distal tubules, alongside intact morphology (HPA tissue IHC: cytoplasmic profile; kidney distal tubules High; standard IHC practice). Strong nuclear or extracellular signal warrants scrutiny because annotated ACSL1 locations are intracellular membranes (UniProt P33121 subcellular location; standard IHC practice). Compare section edges and necrotic areas with well-preserved central tissue, since local damage can distort chromogenic staining (standard IHC practice). Use a no-primary control and peroxidase blocking to investigate signal from the detection workflow, including endogenous enzyme activity (standard IHC practice). Confirm a disputed pattern with an independent antibody or orthogonal expression evidence before assigning it to ACSL1 (standard IHC practice).
Boster reagents
Best ACSL1 / Long-chain-fatty-acid--CoA ligase 1 IHC Antibodies
PB10025 has IHC images from paraffin sections of mouse and rat kidney and human liver cancer tissue, plus an IF/ICC image from U20S cells (catalog image captions).
Real IHC data
Anti-ACSL1 Antibody ®
Cat # PB10025
PB10025 is listed for IHC and IF/ICC, with human, mouse and rat reactivity (catalog applications; catalog reactivity). Its IHC captions show mouse and rat kidney and human liver cancer tissue; its IF caption shows U20S cells (catalog image captions).
Which to pick: For tissue IHC, choose PB10025: its mouse kidney image used a paraffin section, EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody; the fixative is unreported (PB10025 IHC image caption). For IF/ICC, PB10025 has a U20S cell image using 5 μg/ml primary antibody (PB10025 IF image caption). For IHC across species, PB10025 has paraffin-section images from mouse and rat kidney and human liver cancer tissue (PB10025 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P33121 (ACSL1_HUMAN, Long-chain-fatty-acid--CoA ligase 1).
- Human Protein Atlas. ACSL1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ACSL1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. ACSL1 antibody validation summary (3 antibodies).
- Loss of ACSL1 fuels ferroptosis resistance in clear cell renal carcinoma. Cancer biology & therapy 2025 — PMC12505508.
- ACSL1-Dependent Microglial Lipoimmunometabolic Reprogramming Underlies Cognitive Deficits in Alcohol Use Disorder. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13073305.
- The inhibition of Nrf2 accelerates renal lipid deposition through suppressing the ACSL1 expression in obesity-related nephropathy. Renal failure 2019 — PMC6735294.
- ACSL1 Aggravates Thromboinflammation by LPC/LPA Metabolic Axis in Hyperlipidemia Associated Myocardial Ischemia-Reperfusion Injury. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2025 — PMC11923997.
- PubMed PMID:1607358 — UniProt-cited evidence.
- PubMed PMID:8584017 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ACSL1 (Long-chain-fatty-acid--CoA ligase 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB10025). Validated IHC antibody: PB10025 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Appendix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet PB10025); 2. Fixation: Image fixative and duration unreported (datasheet PB10025); 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 PB10025); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB10025); 8. Primary antibody: Rabbit anti-ACSL1, 0.5-1μg/ml (datasheet PB10025); 9. Primary incubation: Overnight at 4 °C (datasheet PB10025). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB10025); Caveat: Adipocyte-rich regions can dominate tissue scoring (HPA tissue IHC); Isoform / epitope: Three isoforms; cytoplasmic epitope coverage is unknown (UniProt); Regulation: Tissue-dependent expression (UniProt). Evidence: UniProt P33121, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet PB10025, PMC13073305, PMC6735294. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ACSL1/immunohistochemistry-ihc-guide.