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ALOX15 / Polyunsaturated fatty acid lipoxygenase ALOX15 · IHC design guide
Design Immunohistochemistry for ALOX15
Plan paraffin-section ALOX15 IHC around cytoplasmic staining in respiratory epithelial cells and subsets of immune cells (HPA tissue IHC). Start with the catalog antibody at 0.5–1 μg/mL (datasheet PB9073) and compare staining in the expected cell populations with controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ALOX15 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 | Cytoplasm in respiratory epithelium and immune subsets (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in respiratory epithelium and immune subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9073) | |
| Positive control | Nasopharynx+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9073) | |
| Caveat | Presumed off-target staining may confound scoring (HPA tissue IHC) | |
| Regulation | IL-13 may redistribute staining (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended ALOX15 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet PB9073) with three published ALOX15 IHC protocols (PMC12289098; PMC9646666; PMC12982294).
| Sample | Paraffin-embedded mouse lung tissue; fixative not specified (datasheet PB9073) |
| Fixation | Image fixative and duration unreported (datasheet PB9073); 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 PB9073); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9073) |
| Primary antibody | Rabbit anti-ALOX15, 0.5-1μg/ml (datasheet PB9073) |
| Primary incubation | Overnight at 4 °C (datasheet PB9073) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9073) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ALOX15-positive staining in basal cells of nasopharynx (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in respiratory epithelial cells and subsets of immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet PB9073); the tumor study used EDTA without reporting its pH (PMC12982294).
Section 2
What Is the Expected ALOX15 Staining Pattern?
ALOX15 staining should be predominantly cytoplasmic in respiratory epithelial cells and subsets of immune cells (HPA tissue IHC: Approved; HPA tissue profile). ALOX15 is predominantly cytosolic, with reported membrane and lipid droplet locations; it has no transmembrane segment (UniProt P16050: subcellular location and topology). Read each cell population separately because staining varies by tissue and cell type (HPA tissue IHC).
| Cytoplasmic staining in nasopharyngeal basal cells or round or early spermatids. | These are strong positive reference patterns: both populations are scored High (HPA tissue IHC). A convincing result follows the cytoplasm and the annotated cell population (HPA tissue profile; UniProt P16050: subcellular location). Compare like cells across sections rather than treating every cell in a positive tissue as positive. |
| Predominantly nuclear staining, or a sharp membrane outline with little cytoplasmic signal. | Treat a nuclear dominant pattern as suspect because ALOX15 is predominantly cytosolic (UniProt P16050: subcellular location). Membrane enrichment is reported under specific conditions, including calcium binding by similarity, so membrane signal alone does not prove an artefact (UniProt P16050: subcellular location). Recheck controls and the cell level pattern. |
| Strong signal in a population recorded as unstained, such as adipocytes in adipose tissue. | Adipocytes are scored Not detected despite tissue enhanced adipose RNA specificity (HPA tissue IHC). Investigate antibody cross reactivity and chromogen produced by endogenous detection activity (general IHC practice). The RNA category does not override the observed cell level protein pattern (HPA tissue IHC). |
| Similar diffuse colour across cells, extracellular space and the negative control. | This does not resolve the cytoplasmic, cell restricted pattern reported for ALOX15 (HPA tissue profile). Review blocking, antibody concentration, washes and endogenous enzyme control as general chromogenic IHC checks (general IHC practice). Interpret the result only after the background is reduced. |
| No staining in a section containing an expected high expressing population. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
💡Expected ALOX15 appearanceCall the IHC result positive when cytoplasmic colour identifies the expected cells, especially High nasopharyngeal basal cells or round or early spermatids (HPA tissue IHC; UniProt P16050: subcellular location); uniform colour in annotated negative cells or across the slide is suspect (HPA tissue IHC; general IHC practice).
| Cell population and reference intensity | HPA scores bone marrow hematopoietic cells, fallopian tube glandular cells, lymph node and tonsil non germinal center cells, and splenic red pulp cells Medium (HPA tissue IHC). Lung macrophages are listed as Low; their faint signal is a less decisive reference than a high expressing population (HPA tissue IHC). |
| Intracellular distribution | ALOX15 is predominantly cytosolic and may become enriched at membranes under specified conditions; lipid droplets are also listed (UniProt P16050: subcellular location). It has no transmembrane segment (UniProt P16050: topology). Score cytoplasmic staining first, while allowing context dependent intracellular variation. |
| Protein forms and antibody epitope | UniProt lists 2 isoforms, a chain spanning residues 2–662, and no signal peptide, propeptide or glycosylation sites (UniProt P16050: isoforms, processing and glycosylation). The supplied records do not identify this antibody's epitope; they cannot predict whether the isoforms stain differently. |
| Strength of the tissue evidence | The HPA tissue profile is Approved, while its reliability note says presumed off target staining was disregarded (HPA tissue IHC: reliability). Three listed antibodies have Approved IHC status, with no ICC status supplied (HPA antibodies). Approval supports using the annotated pattern as a reference; it does not validate every stained structure. |
| What should IF/ICC show? | Use the predominantly cytosolic localisation as the expectation (UniProt P16050: subcellular location). HPA supplies no ICC/IF image, main location or ICC antibody status here, so a cell line specific fluorescent pattern cannot be confirmed from this payload (HPA subcellular; HPA antibodies). The IF/ICC guide handles its own method. |
| Situation | Likely cause | Next action |
|---|---|---|
| A high expressing reference section has no visible chromogen. | The result disagrees with the High HPA scores for nasopharyngeal basal cells and round or early spermatids (HPA tissue IHC); the failing step is undetermined. | Check tissue preservation, antibody preparation, detection reagents and counterstain visibility; review retrieval against the catalog antibody's IHC-P instructions (general IHC practice). Do not infer ALOX15 specific fixation sensitivity from HPA. |
| Signal fills most cells or the space around them. | Diffuse staining obscures HPA's cytoplasmic, cell subset pattern (HPA tissue profile). Excess antibody or insufficient blocking and washing are general IHC possibilities (general IHC practice). | Compare the negative control; optimize blocking, antibody concentration and washes while retaining an expected positive population (general IHC practice). Reassess cell restricted staining after background falls. |
| Brown staining appears in annotated negative adipocytes. | Adipocytes are Not detected in the HPA tissue data (HPA tissue IHC). Cross reactivity or endogenous chromogenic activity is possible, but this observation alone cannot distinguish them (general IHC practice). | Inspect omission and detection controls, apply the appropriate endogenous enzyme control, and compare the suspicious cells with an annotated positive population (general IHC practice; HPA tissue IHC). |
| Only a crisp nuclear pattern is visible. | Nuclear dominance does not match the predominantly cytosolic UniProt location or the HPA cytoplasmic tissue profile (UniProt P16050: subcellular location; HPA tissue profile). | Check the negative control and whether colour lies over nuclei rather than cytoplasm; review counterstain and detection settings (general IHC practice). Avoid scoring nuclear colour alone as ALOX15. |
| Lung macrophages look negative while a strong reference stains. | Lung macrophages are listed as Low, whereas the specified nasopharyngeal basal cells are High (HPA tissue IHC). A weak population can be difficult to judge in one section. | Use the high expressing reference to verify the run, then assess macrophages by cell type and local background (HPA tissue IHC; general IHC practice). Do not use faint macrophage staining as the sole run control. |
| An IF/ICC image appears to contradict the IHC section. | HPA provides no ICC/IF images or defined main subcellular location in the supplied subcellular record (HPA subcellular). UniProt reports context dependent membrane enrichment (UniProt P16050: subcellular location). | Judge the paraffin IHC section against its tissue and cell annotations (HPA tissue IHC). Evaluate the fluorescent result under the separate IF/ICC guide, using its own controls (general IF practice). |
Experiment evidence
IHC & IF images showing ALOX15 staining
PB9073 · IHCIHC analysis of ALOX15 using anti-ALOX15 antibody (PB9073). ALOX15 was detected in a paraffin-embedded section of mouse lung tissue. Heat me
PB9073 · IHCIHC analysis of ALOX15 using anti-ALOX15 antibody (PB9073). ALOX15 was detected in a paraffin-embedded section of rat lung tissue. Heat medi
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000161905-ALOX15/tissue/cerebral+cortex#imid_3483160
HPA · NasopharynxHuman Protein Atlas IHC, Nasopharynx. Source: https://www.proteinatlas.org/ENSG00000161905-ALOX15/tissue/nasopharynx#imid_3483141
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000161905-ALOX15/tissue/colon#imid_3483078
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000161905-ALOX15/tissue/liver#imid_3483079
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ALOX15 IHC & IF
🧪Run nasopharynx first: basal cells should stain (HPA: High in nasopharyngeal basal cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the positive slide, treat adjacent cells without specific staining as internal background, rather than assuming a particular cell type is negative.
Positive control tissue: Nasopharynx (Basal cells, 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 ALOX15; derive a cell-line control from the positive tissue's cell type (Basal cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) section, a matched rabbit IgG isotype control with clonality matched when known, and ALOX15 knockout tissue if available (caption: rabbit primary antibody; standard IHC practice). Check the nasopharynx section for endogenous peroxidase before interpreting DAB staining, and account for endogenous biotin when using the caption’s biotin-based detection system (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9073 paraffin-section caption does not state a fixative (caption: fixative unreported). The reported IHC procedure uses heat-mediated retrieval in EDTA at pH 8.0; whether ALOX15 staining depends on that retrieval condition is unreported (caption: EDTA retrieval). Frozen-section or IF advantages are unreported, and no ALOX15 ICC-IF image-bearing cell lines are supplied (HPA: no ICC-IF images); check nasopharynx sections for background before scoring basal-cell staining (HPA: High in basal cells; standard IHC practice).
HPA tissue IHC evidence for ALOX15
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Presumed off target binding observed and disregarded. Antibody staining in cells/structures not annotated, view images. Protein expression in respiratory epithelia can be correlated with lung RNA-seq 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 |
|---|---|---|---|---|
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Testis | Round or early spermatids | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ALOX15 IHC Tips
Troubleshoot ALOX15 staining in paraffin section chromogenic IHC using the catalog antibody’s mouse lung example and human tissue expression patterns (datasheet PB9073; HPA tissue IHC).
How should I adjust retrieval when ALOX15 staining is weak or uneven?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 (datasheet PB9073). The catalog antibody’s paraffin section mouse lung example used that condition, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet PB9073). If staining remains weak, compare a modestly longer heating interval with the original condition on adjacent sections while holding antibody concentration and detection constant; assess tissue damage and background alongside signal. Avoid treating stronger staining alone as proof of improved specificity: the tissue atlas reports presumed off target binding in its approved ALOX15 IHC assessment (HPA tissue IHC).
Could fixation explain weak ALOX15 staining in my paraffin sections?
Target specific fixation sensitivity is unknown here: the selected paraffin section caption does not state a fixative (datasheet PB9073). Record the fixative, time before fixation, fixation duration, and processing schedule for each specimen, then compare matched sections stained together under the same retrieval and detection conditions. For troubleshooting, start with heat mediated EDTA retrieval at pH 8.0 and the documented 1 μg/ml primary antibody condition before changing multiple variables (datasheet PB9073). Include a tissue control with documented ALOX15 staining, such as nasopharyngeal basal cells, and assess preservation and morphology alongside signal (HPA tissue IHC: High in nasopharyngeal basal cells).
Where should convincing ALOX15 staining appear within positive cells?
Prioritise cytoplasmic staining in morphologically identifiable cells: the tissue atlas describes cytoplasmic expression in respiratory epithelial cells and subsets of immune cells (HPA tissue IHC). ALOX15 is predominantly cytosolic, with membrane enrichment upon calcium binding and reported lipid droplet localisation; its annotated sequence has no transmembrane segment (UniProt P16050 subcellular location and topology). Score a peripheral or punctate pattern only when cell boundaries and staining controls support it, since a brown outline alone does not establish translocation. Check hematoxylin counterstaining to distinguish cytoplasm from adjacent pigment or extracellular deposits. Persistent nuclear only staining warrants a specificity check against the tissue atlas warning about presumed off target binding (HPA tissue IHC).
Can epitope placement explain inconsistent ALOX15 staining across specimens?
First confirm the catalog antibody’s immunogen and mapped epitope before assigning a missing signal to isoform loss; that mapping is not supplied here. The record lists 2 isoforms and a PLAT domain at residues 2–114 followed by a lipoxygenase domain at 115–662 (UniProt P16050). It also lists no signal peptide, transmembrane segment, glycosylation site, or modified residue, which does not establish that any particular epitope survives processing (UniProt P16050 sequence annotations). Compare adjacent sections under identical retrieval and detection conditions, and seek an independently validated antibody to a distinct region if apparent isoform specificity matters to the conclusion.
How can IF help assess a disputed ALOX15 IHC pattern?
Use IF as a separate corroborating experiment and first establish that the chosen antibody works in that application; no IF/ICC images or fixation condition are supplied in the subcellular atlas payload (HPA subcellular). Multiplex ALOX15 with a validated respiratory epithelial or immune cell marker matched to the cells under study, since those compartments are reported in tissue IHC (HPA tissue IHC). Choose a fluorophore in a channel with low measured tissue autofluorescence and include single stain controls for spectral bleed through. For an epitope expected on the cytosolic side, apply and titrate permeabilisation after an IF appropriate fixation step; ALOX15 is predominantly cytosolic and lacks a transmembrane segment (UniProt P16050).
How do I distinguish ALOX15 signal from chromogenic background?
In the documented paraffin section workflow, sections received 10% goat serum block, 1 μg/ml rabbit primary overnight at 4°C, a biotinylated secondary, and DAB development (datasheet PB9073). Add a primary omitted control and, for this biotin based detection system, inspect endogenous biotin and nonspecific secondary staining under the same development time. Quench endogenous peroxidase as a general chromogenic IHC step and compare a shorter DAB development if diffuse brown signal obscures cell boundaries. The atlas specifically notes presumed off target binding, so require reproducible cellular localisation and suitable tissue controls before calling background ALOX15 positive (HPA tissue IHC).
What should I measure when ALOX15 positive cells are sparse? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, then record the percentage of positive cells and staining intensity to calculate an H-score, or count positive cells per mm² of viable tissue. Normalise the positive cell count to the total number of eligible cells in the same compartment when cell composition varies between sections. Report epithelial and immune cell results separately because the atlas describes cytoplasmic expression in respiratory epithelial cells and subsets of immune cells (HPA tissue IHC). Apply one threshold, exposure or scanner setting, and batch control across compared slides; annotate excluded folds, edges, and necrotic areas before analysis.
What would make an apparent ALOX15 positive result unconvincing?
Treat staining as credible when it follows cell morphology and expected cytoplasmic distribution in the scored population; the atlas reports cytoplasmic signal in respiratory epithelial cells and subsets of immune cells (HPA tissue IHC). Recheck a nuclear only pattern against ALOX15’s predominantly cytosolic annotation and absence of a transmembrane segment (UniProt P16050 subcellular location and topology). Discount staining confined to section edges, damaged or necrotic regions, pigment, or sites retaining colour in a primary omitted control; check endogenous peroxidase during DAB troubleshooting. The atlas assessment explicitly notes presumed off target binding, so a positive chromogenic deposit alone is insufficient for a cell type or functional claim (HPA tissue IHC).
Boster reagents
Best ALOX15 / Polyunsaturated fatty acid lipoxygenase ALOX15 IHC Antibodies
PB9073 has real IHC images from paraffin sections of mouse and rat lung and human intestine and mammary cancer tissue (PB9073 image captions). No IF image is supplied (catalog: IF images absent).
Real IHC data
Anti-15 Lipoxygenase 1/ALOX15 Antibody ®
Cat # PB9073
PB9073 is the only card and is listed for IHC in human, mouse and rat (catalog: PB9073 applications and reactivity). Its IHC captions show paraffin sections of mouse and rat lung and human intestine and mammary cancer tissue (PB9073 image captions).
Which to pick: For tissue IHC, choose PB9073: its paraffin section captions document EDTA pH 8.0 retrieval and 1 μg/ml primary antibody (PB9073 image captions); the fixative is unreported (PB9073 image captions). For cross species IHC, PB9073 is listed for human, mouse and rat (catalog: PB9073 reactivity). No IF/ICC validated SKU is provided here (catalog: PB9073 applications and IF images absent).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P16050 (LOX15_HUMAN, Polyunsaturated fatty acid lipoxygenase ALOX15).
- Human Protein Atlas. ALOX15 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ALOX15 subcellular location (ICC-IF): Highest expression in MCF-7: 20.8 nTPM.
- Human Protein Atlas. ALOX15 antibody validation summary (3 antibodies).
- Predictive significance of 15(S)-hydroxyeicosatetraenoic acid for eosinophilic chronic rhinosinusitis with nasal polyps. Asia Pacific allergy 2025 — PMC12289098.
- Non-invasive biomarkers of eosinophilic esophagitis. Acta bio-medica : Atenei Parmensis 2021 — PMC9431887.
- Identification of a Feed-Forward Loop Between 15(S)-HETE and PGE2 in Human Amnion at Parturition. Journal of lipid research 2022 — PMC9646666.
- ALOX15 exerts tumor-suppressive role in head and neck squamous cell carcinoma via ERK pathway. Functional & integrative genomics 2026 — PMC12982294.
- PubMed PMID:3202857 — UniProt-cited evidence.
- PubMed PMID:9224951 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ALOX15 (Polyunsaturated fatty acid lipoxygenase ALOX15). Expected localisation Cytoplasm in respiratory epithelium and immune subsets (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9073). Validated IHC antibody: PB9073 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Nasopharynx (Basal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse lung tissue; fixative not specified (datasheet PB9073); 2. Fixation: Image fixative and duration unreported (datasheet PB9073); 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 PB9073); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9073); 8. Primary antibody: Rabbit anti-ALOX15, 0.5-1μg/ml (datasheet PB9073); 9. Primary incubation: Overnight at 4 °C (datasheet PB9073). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9073); Caveat: Presumed off-target staining may confound scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: IL-13 may redistribute staining (UniProt). Evidence: UniProt P16050, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in MCF-7: 20.8 nTPM, Datasheet PB9073, PMC12289098, PMC9646666. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ALOX15/immunohistochemistry-ihc-guide.