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SMOX / Spermine oxidase · IHC design guide
Design Immunohistochemistry for SMOX
Plan chromogenic SMOX IHC in paraffin sections using the catalog antibody (datasheet A05519-2). Assess cytoplasmic and nuclear staining, with Leydig cells as a high-staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SMOX 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 and nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining; Leydig cells high (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05519-2) | |
| Positive control | Testis+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 A05519-2) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Stimulus-specific regulation unreported (UniProt) | |
| Isoform / epitope | Six isoforms; epitope coverage across isoforms is unknown (UniProt) |
Section 1
Recommended SMOX IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A05519-2). The published IHC protocols below describe paraffin tissues, mouse oral mucosa, and lung cancer sections (PMC4345146; PMC9414028; PMC8339854).
| Sample | Paraffin-embedded human diffuse large B cell lymphoma tissue; fixative not specified (datasheet A05519-2) |
| Fixation | Image fixative and duration unreported (datasheet A05519-2); 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 A05519-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05519-2) |
| Primary antibody | Rabbit anti-SMOX, 2-5 μg/ml (datasheet A05519-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05519-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05519-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SMOX-positive staining in leydig cells of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A05519-2). Use citrate pH 6.0 when reproducing the mouse oral study (PMC9414028).
Section 2
What Is the Expected SMOX Staining Pattern?
SMOX is reported in the cytoplasm and nucleus (UniProt Q9NWM0; HPA tissue IHC). In paraffin sections, expect staining across many tissues, with particularly strong staining in testicular Leydig cells and medium staining in several listed cell populations (HPA tissue IHC). HPA rates its tissue IHC pattern Approved, with medium consistency between staining and RNA data (HPA tissue IHC). SMOX has no transmembrane segment (UniProt Q9NWM0 topology).
| Leydig cells show strong nuclear and/or cytoplasmic staining, while nearby structures remain distinguishable. | This is a useful positive reference: HPA reports High staining in Leydig cells and a cytoplasmic and nuclear pattern across most tissues (HPA tissue IHC). Assess the expected cells and compartments together; strong color alone does not establish specificity (standard IHC practice). |
| Signal is confined to a sharp cell-surface rim or extracellular material, with no nuclear or cytoplasmic staining. | That distribution does not match the reported tissue pattern (HPA tissue IHC) or the absence of a transmembrane segment (UniProt Q9NWM0 topology). Review morphology and controls before interpreting it as SMOX; HPA ICC-IF also reports a nuclear-membrane location, which should not be confused with a cell-surface rim (HPA subcellular ICC-IF). |
| An unexpected cell population stains more strongly than the documented positive cells. | Compare against the cell-specific references: Leydig cells are High; adipocytes, selected glandular and respiratory epithelial cells, glia, and cerebellar granular-layer cells are Medium (HPA tissue IHC). Unexpected staining warrants a cross-reactivity check, but broad distribution alone is insufficient evidence of artefact because SMOX is widely expressed (UniProt Q9NWM0). |
| Color spreads across cells and tissue spaces, obscuring cell borders and nuclei. | Treat this as background until controls show otherwise (standard IHC practice). HPA describes cellular cytoplasmic and nuclear expression, not an uninterpretable tissue-wide haze (HPA tissue IHC). Compare a no-primary control and inspect whether the chromogen is restricted to identifiable cells (standard IHC practice). |
| The known-positive Leydig cells show no convincing signal. | HPA reports High staining in these cells, making an entirely blank result a reason to review assay performance before scoring other tissues negative (HPA tissue IHC). Check the positive control, antibody application, retrieval, and detection steps as general IHC troubleshooting; the supplied sources do not establish an SMOX-specific retrieval or fixation requirement. |
💡Expected SMOX appearanceA convincing positive shows identifiable nuclear and/or cytoplasmic staining, strongest in Leydig cells (High) and potentially weaker in HPA-listed Medium cells (HPA tissue IHC); an isolated cell-surface rim or diffuse tissue-wide haze is suspect against that reference pattern (HPA tissue IHC; UniProt Q9NWM0 topology).
| Tissue and cell reference | Use the named cell population when judging intensity: Leydig cells are High; adipocytes, adrenal, appendix and breast glandular cells, bronchial respiratory epithelial cells, caudate glia, and cerebellar granular-layer cells are Medium (HPA tissue IHC). Cervical glandular, smooth-muscle, and bone-marrow hematopoietic cells are listed Low, not absent (HPA tissue IHC). |
| Compartment | Tissue IHC reports cytoplasmic and nuclear expression in most tissues (HPA tissue IHC), consistent with the UniProt cytoplasm and nucleus annotation (UniProt Q9NWM0). A nucleus-only or cytoplasm-only view of an individual cell should be assessed against tissue morphology and controls before being called incorrect; the sources do not prescribe a fixed nuclear-to-cytoplasmic ratio. |
| Antibody evidence | HPA047117 is Approved for IHC, and HPA describes medium consistency between antibody staining and RNA expression (HPA antibodies; HPA tissue IHC). This supports using the published pattern as a reference, while leaving room to investigate discordant staining; the supplied IHC record does not claim the Enhanced validation category (HPA antibodies). |
| Isoforms and protein features | UniProt lists 6 SMOX isoforms, a single chain spanning residues 1–555, no signal peptide or propeptide, no annotated glycosylation sites, and no transmembrane segment (UniProt Q9NWM0). The payload does not identify the antibody epitope, so these features cannot establish which isoforms it detects or predict an SMOX-specific retrieval response. |
| Does ICC-IF define the paraffin IHC pattern? | HPA ICC-IF places SMOX mainly in the nucleoplasm, with additional nuclear-membrane, vesicle, and cytosol locations; images are listed for A-549, SiHa, and U2OS (HPA subcellular ICC-IF). This is useful compartment context, but the ICC-Supported antibody HPA060198 differs from the IHC-Approved antibody HPA047117 (HPA antibodies). Interpret paraffin staining against the tissue IHC record. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control Leydig cells are blank or barely visible. | The run may have failed, or the chosen conditions may provide inadequate detection; High Leydig-cell staining is the HPA reference (HPA tissue IHC). | First verify the positive-control section, antibody and detection reagents, and that the retrieval and staining steps ran as intended (standard IHC practice). Do not infer an SMOX-specific retrieval or fixation sensitivity from this result. |
| All tissue structures, including spaces without identifiable cells, develop color. | Nonspecific background or endogenous detection activity can obscure cellular staining (standard IHC practice). | Review the no-primary control, blocking, washing, and chromogen development (standard IHC practice). If using an HRP detection system, check the endogenous-peroxidase control (standard IHC practice); retain only interpretable nuclear or cytoplasmic cellular signal (HPA tissue IHC). |
| Only a sharp cell-surface outline stains. | A surface-only pattern conflicts with the reported cytoplasmic and nuclear tissue staining (HPA tissue IHC) and SMOX lacking a transmembrane segment (UniProt Q9NWM0 topology). | Check morphology and no-primary staining, then compare with a documented positive section (standard IHC practice; HPA tissue IHC). Keep the ICC-IF nuclear-membrane observation distinct from a plasma-membrane outline (HPA subcellular ICC-IF). |
| Unexpected cells stain while the documented positive population also stains. | SMOX is widely expressed (UniProt Q9NWM0), so location alone cannot classify the extra signal; cross-reactivity remains possible (standard IHC practice). | Compare cell identity, compartment, and intensity with HPA's named tissue observations, then inspect controls (HPA tissue IHC; standard IHC practice). Report unexpected staining as uncertain if those checks cannot resolve it. |
| Low-staining tissue is scored as definitively SMOX-negative. | HPA lists cervical glandular cells, smooth-muscle cells, and bone-marrow hematopoietic cells as Low, while listing no negative tissue examples in this payload (HPA tissue IHC). | Record weak versus absent signal explicitly and judge the section alongside a positive control (standard IHC practice; HPA tissue IHC). Avoid designating an HPA Low population as a validated negative control. |
| An ICC-IF localization image appears inconsistent with the paraffin section. | The records use different modalities and antibodies: HPA060198 is ICC Supported, whereas HPA047117 is IHC Approved (HPA antibodies). | Use the tissue IHC pattern for the paraffin-section decision and the ICC-IF record as compartment context (HPA tissue IHC; HPA subcellular ICC-IF). Review the paraffin controls before calling a discrepancy biological (standard IHC practice). |
Experiment evidence
IHC & IF images showing SMOX staining
A05519-2 · IHCIHC analysis of SMOX using anti-SMOX antibody (A05519-2). SMOX was detected in a paraffin-embedded section of human diffuse large B cell lym
A05519-2 · IHCIHC analysis of SMOX using anti-SMOX antibody (A05519-2). SMOX was detected in a paraffin-embedded section of human duodenal papilla adenoca
A05519-2 · IFIF analysis of SMOX using anti-SMOX antibody (A05519-2) and anti-Beta Tubulin antibody (M01857-3). SMOX was detected in immunocytochemical s
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000088826-SMOX/tissue/cerebral+cortex#imid_12878735
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000088826-SMOX/tissue/colon#imid_12878645
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000088826-SMOX/tissue/liver#imid_12878654
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000088826-SMOX/tissue/kidney#imid_12878655
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA060198; green = SMOX, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA060198; green = SMOX, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SMOX IHC & IF
🧪Run testis first and look for SMOX staining in Leydig cells (HPA: High in Leydig cells). HPA detects SMOX in all 45 scored tissues, so use no-primary and isotype controls for the negative; cells on the testis slide without specific staining should show only background chromogen and are not validated SMOX-negative cells (HPA: no negative tissue listed).
Positive control tissue: Testis (Leydig cells, HPA High)
Negative control tissue: None in HPA: SMOX is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SMOX in A-549, SiHa, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a rabbit IgG isotype control matched to the primary antibody’s class and clonality, and SMOX-knockout tissue if available as a biological negative (selected A05519-2 caption: rabbit primary and anti-rabbit secondary). For HRP/DAB detection, quench endogenous peroxidase and check the testis control section for residual background (selected A05519-2 caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative is unreported in the selected A05519-2 paraffin-section caption. That caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required (selected A05519-2 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; endogenous peroxidase background should be checked in the testis section when using HRP/DAB (selected A05519-2 caption: HRP/DAB detection).
HPA tissue IHC evidence for SMOX
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SMOX is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SMOX IHC Tips
Troubleshoot SMOX staining in paraffin sections by checking retrieval, cell compartment, controls, and scoring against the reported IHC pattern.
Which retrieval condition should I try first for weak SMOX staining?
Start with heat mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet A05519-2). The selected tissue image used this condition before incubation with 2 μg/ml catalog antibody overnight at 4°C (caption A05519-2). If staining is weak, adjust heating duration within your instrument's validated range and compare adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Record tissue integrity alongside signal, because excessive heating can damage morphology and make cytoplasmic or nuclear staining difficult to judge (standard IHC practice; HPA tissue IHC: cytoplasmic and nuclear expression).
Could fixation explain weak or uneven SMOX IHC staining?
The selected paraffin section caption does not report a fixative, so SMOX specific fixation sensitivity is unknown (caption A05519-2). Review fixation duration and section history, then compare specimens processed under matched conditions before attributing intensity differences to biology (standard IHC practice). For a processing comparison, stain adjacent sections with the same EDTA pH 8.0 retrieval and 2 μg/ml antibody condition used in the selected image (caption A05519-2). Preserve a morphology check and a no primary control in each run, since damaged tissue and detection background can mimic a fixation effect (standard IHC practice).
How should I judge nuclear versus cytoplasmic SMOX staining?
Evaluate cytoplasmic and nuclear DAB signal separately, because both compartments are reported for SMOX (UniProt Q9NWM0: cytoplasm and nucleus; HPA tissue IHC: cytoplasmic and nuclear expression). A nucleoplasmic emphasis is compatible with the supported subcellular IF location, but that observation does not establish the expected intensity ratio in paraffin IHC (HPA subcellular: nucleoplasm supported). Check that a nuclear call follows intact nuclei across cells rather than section folds or counterstain variation (standard IHC practice). Score the two compartments independently across comparable cell populations, using adjacent negative controls to assess diffuse DAB haze (standard IHC practice).
Can this IHC result distinguish SMOX isoforms?
Do not assign an IHC staining pattern to a particular splice isoform without epitope mapping and isoform specific validation (UniProt Q9NWM0: 6 isoforms; supplied antibody evidence: no epitope mapping). SMOX is annotated as a 555 amino acid chain without a signal peptide, propeptide, transmembrane segment, or annotated glycosylation sites (UniProt Q9NWM0). Those annotations alone do not show which isoforms the catalog antibody recognizes in fixed sections (UniProt Q9NWM0; supplied antibody evidence: no epitope mapping). If an isoform claim matters, establish antibody reactivity with suitable isoform controls and compare staining under identical retrieval and detection conditions (standard IHC validation practice).
How can IF help resolve ambiguous SMOX IHC staining?
Use IF as a separate localisation check, pairing SMOX with a validated marker for the cell population under study, such as Leydig cells when examining testis (HPA tissue IHC: high in Leydig cells). Select spectrally separated fluorophores and inspect an unstained section for autofluorescence before assigning faint signal to SMOX (standard IF practice). For access to cytoplasmic or nuclear epitopes, test controlled permeabilisation after fixation; SMOX has no annotated transmembrane segment, while its reported locations include both compartments (UniProt Q9NWM0). Include single stain and no primary controls so bleed through and background can be separated from colocalisation (standard IF practice).
What should I check when SMOX DAB staining is diffuse?
Inspect a no primary control and block endogenous peroxidase before DAB development to identify enzyme driven background (standard chromogenic IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a peroxidase conjugated secondary, and DAB (caption A05519-2). If controls remain clean but staining is diffuse, compare antibody concentration and wash stringency systematically while keeping retrieval and exposure conditions matched (standard IHC practice). Judge improvement by preserved cellular boundaries and the reported cytoplasmic or nuclear pattern, rather than by lower overall brown intensity alone (HPA tissue IHC: cytoplasmic and nuclear expression; standard IHC practice).
How should I quantify SMOX across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear and cytoplasmic compartments separately, since both are reported in tissue IHC (HPA tissue IHC: cytoplasmic and nuclear expression). For each compartment, record percent positive viable cells and intensity, or calculate an H score from intensity categories 0–3, yielding 0–300 (standard IHC scoring practice). If cell abundance varies, also report positive cells per mm² and normalize counts to viable tissue area or the relevant cell population (standard IHC practice). Keep retrieval, DAB development, imaging, thresholds, and control sections consistent across specimens before comparing scores (standard IHC practice).
When is apparent SMOX positivity likely to be artefactual?
Treat reproducible cytoplasmic or nuclear staining in intact cells as more plausible than signal confined to tissue edges, folds, or necrosis (UniProt Q9NWM0: cytoplasm and nucleus; standard IHC practice). Compare cell identity with the expected tissue context: Leydig cells show high staining in testis, while hematopoietic cells in bone marrow are reported low (HPA tissue IHC). Brown signal in a no primary control suggests detection background, including endogenous peroxidase, rather than antibody dependent SMOX staining (standard chromogenic IHC practice). Interpret modest differences cautiously because the tissue IHC profile is rated Approved with medium consistency against RNA expression (HPA tissue IHC).
Boster reagents
Best SMOX / Spermine oxidase IHC Antibodies
The IHC-validated anti-SMOX antibody has paraffin-section images from human tumors and an IF/ICC image from A549 cells; listed reactivity includes Human, Mouse, and Rat (catalog IHC/IF captions; catalog reactivity).
Real IHC data
Anti-SMOX Antibody ®
Cat # A05519-2
A05519-2 has IHC images from human paraffin sections of diffuse large B cell lymphoma, duodenal papilla adenocarcinoma, endometrioid adenocarcinoma, and glioblastoma (catalog IHC captions). The same SKU has an IF/ICC image in A549 cells and lists Human, Mouse, and Rat reactivity (catalog IF caption; catalog reactivity).
Which to pick: For tissue IHC, choose A05519-2: its own caption shows a human lymphoma paraffin section with EDTA retrieval at pH 8.0; the fixative is unreported (A05519-2 IHC caption). For IF/ICC, the same SKU has an A549-cell IF image and lists both applications (A05519-2 IF caption; catalog applications). For cross-species work, A05519-2 lists Human, Mouse, and Rat reactivity, while the supplied IHC images are human and the IF image is A549; clonality is unreported (catalog reactivity; A05519-2 IHC/IF captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NWM0 (SMOX_HUMAN, Spermine oxidase).
- Human Protein Atlas. SMOX tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SMOX subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear membrane, vesicles and cytosol..
- Human Protein Atlas. SMOX antibody validation summary (2 antibodies).
- Increased Helicobacter pylori-associated gastric cancer risk in the Andean region of Colombia is mediated by spermine oxidase. Oncogene 2015 — PMC4345146.
- Activation of EGFR and ERBB2 by Helicobacter pylori results in survival of gastric epithelial cells with DNA damage. Gastroenterology 2014 — PMC4035375.
- Microbiome dysbiosis inhibits carcinogen-induced murine oral tumorigenesis. Journal of Cancer 2022 — PMC9414028.
- SMOX expression predicts the prognosis of non-small cell lung cancer. Annals of translational medicine 2021 — PMC8339854.
- PubMed PMID:11454677 — UniProt-cited evidence.
- PubMed PMID:12398765 — UniProt-cited evidence.
- PubMed PMID:18422650 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SMOX (Spermine oxidase). Expected localisation Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05519-2). Validated IHC antibody: A05519-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Leydig cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human diffuse large B cell lymphoma tissue; fixative not specified (datasheet A05519-2); 2. Fixation: Image fixative and duration unreported (datasheet A05519-2); 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 A05519-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05519-2); 8. Primary antibody: Rabbit anti-SMOX, 2-5 μg/ml (datasheet A05519-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05519-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A05519-2); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: Six isoforms; epitope coverage across isoforms is unknown (UniProt); Regulation: Stimulus-specific regulation unreported (UniProt). Evidence: UniProt Q9NWM0, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05519-2, PMC4345146, PMC9414028. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SMOX/immunohistochemistry-ihc-guide.