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SELENBP1 / Methanethiol oxidase · IHC design guide
Design Immunohistochemistry for SELENBP1
Plan paraffin-section SELENBP1 IHC with the catalog antibody at 2–5 μg/mL (datasheet A04444-3). Score cytoplasmic staining and note occasional nuclear signal; liver hepatocytes are a high-staining reference, while adrenal glandular cells are not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SELENBP1 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 | Predominantly cytoplasmic; nuclear in some cases (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in glandular cells and hepatocytes; sometimes nuclear (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04444-3) | |
| Positive control | Colon+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Lung, colorectal and ovarian tumors may stain less (UniProt) | |
| Regulation | Down in lung/colorectal/ovarian cancer (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope impact is undetermined (UniProt) |
Section 1
Recommended SELENBP1 IHC & IF Protocols
The catalog antibody specifies EDTA pH 8.0 retrieval (datasheet: A04444-3). Published IHC protocols below cover human and mouse tissues, head and neck cancer, and colorectal cancer (PMC5828193; PMC5008557; PMC9438969).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet A04444-3) |
| Fixation | Image fixative and duration unreported (datasheet A04444-3); 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 A04444-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04444-3) |
| Primary antibody | Rabbit anti-SELENBP1, 2-5 μg/ml (datasheet A04444-3) |
| Primary incubation | Overnight at 4 °C (datasheet A04444-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04444-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SELENBP1-positive staining in endocrine cells of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with high levels in most glandular epithelia and in hepatocytes. Nuclear expression in several cases. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A04444-3); citrate pH 6 for 20 minutes is a published alternative (PMC5008557: IHC methods).
Section 2
What Is the Expected SELENBP1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic SELENBP1 staining in glandular epithelia and hepatocytes, with nuclear staining in some cases (HPA: tissue IHC, Enhanced reliability). High staining is documented in liver hepatocytes, intestinal Paneth cells and lung alveolar type II cells (HPA: High). UniProt also places SELENBP1 in the cytosol and nucleus and reports no transmembrane segment (UniProt Q13228: subcellular location and topology).
| Strong cytoplasmic staining in hepatocytes, intestinal Paneth cells or lung alveolar type II cells. | This matches documented high staining in those cells (HPA: High). Compare the signal with nearby structures on the same section: a cell-specific cytoplasmic pattern is more persuasive than a uniformly colored field (general IHC practice). |
| Signal appears only as a sharp membrane rim, or only in a compartment inconsistent with the tissue pattern. | Recheck interpretation and staining specificity. UniProt reports cytosolic and nuclear localization with no transmembrane segment (UniProt Q13228: location and topology); HPA tissue IHC chiefly shows cytoplasm, with nuclear staining in several cases (HPA: tissue IHC). A nuclear signal alone is therefore not automatically an artefact. |
| The strongest color is in an unexpected cell population rather than the documented positive cells. | Consider cross-reactivity or detection background before calling it SELENBP1 (general IHC practice). Check cell identity against the section morphology and HPA's cell-level observations; HPA reports high staining in hepatocytes and intestinal Paneth cells, while some other cell populations are not detected (HPA: tissue IHC). |
| Pale, diffuse color covers positive cells, negative cells and surrounding tissue. | A field-wide haze cannot establish cell-level SELENBP1 expression (general IHC practice). The HPA pattern is predominantly cytoplasmic and varies among cell populations (HPA: tissue IHC); assess background with the detection control and compare areas expected to stain differently. |
| No staining appears in an included liver hepatocyte or intestinal Paneth-cell control. | Treat the run as uninformative until the positive control works: both populations have high documented staining (HPA: High). Review control identity, antibody and detection setup, and section quality before interpreting a study specimen as negative (general IHC practice). |
💡Expected SELENBP1 appearanceCall a result positive when identifiable hepatocytes or intestinal Paneth cells show clear, strong cytoplasmic staining (HPA: High; HPA: cytoplasmic profile); color spread evenly across unrelated cells or tissue is a background warning (general IHC practice).
| Tissue and cell selection | HPA records high staining in liver hepatocytes, duodenal and small-intestinal Paneth cells, and lung alveolar type II cells (HPA: High). It also records not-detected glandular cells in adrenal gland and cervix (HPA: Not detected). Choose controls by cell population, not tissue name alone. |
| Compartment used for scoring | Predominantly cytoplasmic staining, with nuclear expression in several cases, is the tissue-IHC reference (HPA: tissue IHC). Cytosolic and nuclear localization agrees with UniProt, which lists no transmembrane segment (UniProt Q13228: location and topology). Record compartments separately when scoring. |
| Antibody validation | HPA lists IHC validation as Enhanced for HPA005741, HPA011731 and CAB008366 (HPA: antibodies). That supports the reference tissue pattern; it does not establish that every antibody will give the same staining under every assay condition. |
| Isoform recognition | UniProt lists four SELENBP1 isoforms (UniProt Q13228: isoforms 1–4). The supplied record does not map antibody epitopes to isoforms, so a different pattern cannot be assigned to one isoform from these data alone. |
| IHC versus ICC-IF readout | HPA tissue IHC describes chiefly cytoplasmic staining with nuclear expression in some cases (HPA: tissue IHC), whereas HPA ICC-IF reports supported nucleolar localization (HPA: subcellular). Keep these assay-specific observations distinct when interpreting an image. |
| Situation | Likely cause | Next action |
|---|---|---|
| The study section is negative, and the positive control is also blank. | The assay may not have demonstrated detectable staining; high staining is expected in liver hepatocytes or intestinal Paneth cells (HPA: High). | Confirm the control contains the named cells, then check antibody application, detection reagents and counterstain balance before judging the study section (general IHC practice). |
| A purported negative control shows strong cell-specific staining. | The control may contain another expressing cell type, or the signal may be nonspecific. HPA's not-detected calls apply to named cells, such as adrenal glandular cells, rather than every cell in the tissue (HPA: Not detected). | Identify the stained cells morphologically; compare the exact cell population with HPA's annotation and review detection controls before rejecting the control (general IHC practice). |
| Color appears throughout the section without a clear cellular pattern. | Diffuse background can obscure the predominantly cytoplasmic reference pattern (HPA: tissue IHC); its cause cannot be assigned from the appearance alone. | Inspect the no-primary control, wash and blocking steps, and chromogen development as general IHC checks. Reassess the cells only after background is controlled. |
| Signal follows tissue edges, debris or many unrelated cells. | This distribution is less consistent with the cell-level HPA observations (HPA: tissue IHC) and may reflect nonspecific staining or detection activity (general IHC practice). | Review section morphology and the no-primary control. If using an enzyme-based chromogen, verify the relevant endogenous activity control before assigning SELENBP1 staining (general IHC practice). |
| The slide shows nuclear staining as well as cytoplasm. | Nuclear expression occurs in several tissue-IHC cases (HPA: tissue IHC), and UniProt lists nuclear localization (UniProt Q13228: subcellular location). | Score nuclear and cytoplasmic signal separately and compare their cell distribution with the positive control; do not discard a nuclear component solely because the dominant tissue pattern is cytoplasmic. |
| Does nucleolar staining in an IF or ICC image contradict the paraffin-section IHC result? | HPA reports supported nucleolar localization in ICC-IF (HPA: subcellular), while its tissue-IHC profile is chiefly cytoplasmic with nuclear staining in several cases (HPA: tissue IHC). | Interpret each image against its own assay reference and cell identity. Use the separate IF/ICC guide for that assay; the tissue-IHC observations alone cannot resolve an IF localization discrepancy. |
Experiment evidence
IHC & IF images showing SELENBP1 staining
A04444-3 · IHCIHC analysis of SBP/SELENBP1 using anti-SBP/SELENBP1 antibody (A04444-3). SBP/SELENBP1 was detected in a paraffin-embedded section of human
A04444-3 · IHCIHC analysis of SBP/SELENBP1 using anti-SBP/SELENBP1 antibody (A04444-3). SBP/SELENBP1 was detected in a paraffin-embedded section of human
A04444-3 · IFIF analysis of SBP/SELENBP1 using anti-SBP/SELENBP1 antibody (A04444-3). SBP/SELENBP1 was detected in a paraffin-embedded section of human r
PA2216 · IHCAnti-SBP/SELENBP1 antibody, PA2216, IHC(P) IHC(P): Human Liver Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000143416-SELENBP1/tissue/cerebral+cortex#imid_3060944
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000143416-SELENBP1/tissue/colon#imid_3060862
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000143416-SELENBP1/tissue/liver#imid_3060871
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000143416-SELENBP1/tissue/kidney#imid_3060864
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA011731; green = SELENBP1, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA011731; green = SELENBP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SELENBP1 IHC & IF
🧪Run liver first: hepatocytes should stain strongly (HPA: High in hepatocytes). Use adrenal gland as the negative tissue, focusing on glandular cells (HPA: Not detected in glandular cells); on the liver slide, non-hepatocyte cells should show only background if used as internal negative comparators, but their SELENBP1 status is not established by the supplied HPA row.
Positive control tissue: Colon (Endocrine cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SELENBP1 in HeLa, U-251MG, U2OS, with annotated localisation: Nucleoli (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (catalog antibody caption: rabbit primary; standard IHC practice). Use SELENBP1 knockout tissue or an antigen-peptide blocking control if available, and quench endogenous peroxidase before HRP/DAB detection in liver sections (standard IHC practice; catalog antibody caption: HRP/DAB).
⚠️Feasibility: The selected A04444-3 paraffin-section caption does not state a fixative, and no target-specific fixation window or fixation effect is reported (selected-SKU tissue-IHC caption). Heat retrieval in EDTA at pH 8.0 is a documented starting condition, but the caption does not establish that retrieval is required (selected-SKU tissue-IHC caption). The supplied evidence does not show that frozen sections or IF are easier than paraffin IHC; check liver sections for residual endogenous peroxidase signal (selected-SKU tissue-IHC caption: paraffin HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for SELENBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Paneth cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular 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 SELENBP1 IHC Tips
Troubleshoot SELENBP1 staining in paraffin section chromogenic IHC using the catalog antibody’s tissue example and reported expression patterns.
What retrieval should I try when SELENBP1 staining is weak in paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 for SELENBP1 paraffin section IHC (datasheet A04444-3). The catalog image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody, so keep those conditions together during initial troubleshooting (datasheet A04444-3). If staining remains weak, compare a small retrieval time series on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Include a positive hepatocyte or glandular epithelial region and inspect morphology, because excessive heating can damage sections and make an apparent signal gain difficult to interpret (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or patchy SELENBP1 staining?
The selected tissue example identifies a paraffin section but does not state its fixative, so SELENBP1 sensitivity to fixation is unknown for this antibody (datasheet A04444-3). Record each specimen’s fixative and fixation duration, then compare similarly processed sections using the same EDTA pH 8.0 retrieval and detection conditions (datasheet A04444-3; standard IHC practice). Uneven fixation or processing can produce patchy immunostaining, so examine whether staining follows section thickness or tissue edges before assigning it to SELENBP1 expression (standard IHC practice). Use an internal positive cell population where available, and avoid inferring fixation tolerance from reported tissue expression or protein topology (HPA tissue IHC; UniProt Q13228 topology; standard IHC practice).
Should SELENBP1 appear in the cytoplasm, nucleus, or membrane by IHC?
Expect predominantly cytoplasmic staining in many glandular epithelia and hepatocytes, while allowing nuclear staining in some settings (HPA tissue IHC). UniProt lists cytosol and nucleus and notes possible Golgi and autophagosome membrane association by similarity; SELENBP1 has no annotated transmembrane segment (UniProt Q13228 subcellular location and topology). Score cytoplasmic and nuclear compartments separately rather than treating either pattern alone as proof of antibody specificity (HPA tissue IHC; standard IHC practice). If only crisp surface membrane staining appears, check a positive control and secondary only control before interpretation, because that pattern does not follow the reported predominant tissue distribution (HPA tissue IHC; standard IHC practice).
How should I troubleshoot discordant staining if the antibody epitope is unknown?
SELENBP1 has 4 annotated isoforms, but the supplied antibody example does not identify its epitope or establish isoform coverage (UniProt Q13228 isoforms; datasheet A04444-3). Check the antibody’s available immunogen information before attributing regional staining differences to alternative splicing, and compare sections processed in the same staining run (standard IHC practice). UniProt reports acetylation at residue 2 and phosphorylation at residues 111, 371, and 467; their effect on antibody binding is unreported here (UniProt Q13228 modified residues). If staining disagrees with expected positive cells, use an independently validated epitope or orthogonal expression measure where available rather than assigning an isoform specific pattern (HPA tissue IHC; standard IHC practice).
How can IF help resolve ambiguous SELENBP1 IHC localisation?
Use IF as a separate validation experiment and multiplex SELENBP1 with a marker for the expected cell population, such as hepatocytes, to check which cells carry the signal (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, and reserve a spectrally distinct channel for the cell marker (standard IF practice). For a cytosolic or nuclear epitope, test gentle permeabilisation after fixation; an accessible surface epitope would require a different approach, but SELENBP1 has no annotated transmembrane segment (UniProt Q13228 topology; standard IF practice). HPA reports supported nucleolar localisation in ICC/IF, while tissue IHC also reports cytoplasmic staining, so compare compartments without assuming both preparations must look identical (HPA subcellular; HPA tissue IHC).
What should I adjust when DAB background obscures SELENBP1 staining?
The catalog tissue example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, an HRP secondary for 30 minutes at 37°C, and DAB (datasheet A04444-3). If diffuse background dominates, titrate the primary concentration and shorten chromogen development while retaining a positive control on the same run (standard IHC practice). Include a primary omitted control, block endogenous peroxidase, and inspect endogenous pigment before calling brown deposits specific (standard IHC practice). Compare background in reported low or undetected cell populations with signal in positive cells; uneven edge staining also warrants review of section handling and reagent coverage (HPA tissue IHC; standard IHC practice).
How should I quantify SELENBP1 IHC across specimens with mixed cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: HPA reports strong staining in hepatocytes and several epithelial cell types, with nuclear expression in some cases (HPA tissue IHC). For chromogenic sections, report an H-score from intensity and percentage of positive target cells, or report percentage positive when intensity cannot be graded reliably (standard IHC practice). For spatial questions, count positive cells per mm² of viable, annotated tissue and normalise to the number of eligible cells or measured area (standard IHC practice). Apply identical imaging, thresholds, retrieval, and DAB development across specimens, and score cytoplasmic and nuclear signals separately when both occur (HPA tissue IHC; standard IHC practice).
How can I distinguish real SELENBP1 signal from staining artefacts?
Look for reproducible cytoplasmic staining in expected cells, such as hepatocytes or colon endocrine cells, while recognising that nuclear staining can also occur (HPA tissue IHC). Strong isolated surface membrane staining needs scrutiny because SELENBP1 lacks a transmembrane segment and reported tissue staining is chiefly cytoplasmic (UniProt Q13228 topology; HPA tissue IHC). Exclude tissue edges, folds, necrotic areas, and uneven DAB deposits from scoring, and check a primary omitted control for endogenous enzyme or detection background (standard IHC practice). Interpret absent staining cautiously in a single section: compare a documented positive region and confirm that retrieval and detection worked in the same run (HPA tissue IHC; datasheet A04444-3; standard IHC practice).
Boster reagents
Best SELENBP1 / Methanethiol oxidase IHC Antibodies
IHC-validated SELENBP1 antibodies have human tissue images; one also has human rectal cancer IF images and listed human, mouse and rat reactivity (catalog IHC/IF captions; catalog reactivity).
Real IHC data
Anti-SBP/SELENBP1 Antibody ®
Cat # A04444-3
Real IHC data
Anti-SBP/SELENBP1 Antibody ®
Cat # PA2216
A04444-3 has IHC images from paraffin sections of human spleen, breast cancer, colon adenocarcinoma and prostate adenocarcinoma, plus an IF image from paraffin-section human rectal cancer (A04444-3 image captions). PA2216 has a human liver cancer IHC(P) image and lists human reactivity (PA2216 IHC caption; catalog reactivity).
Which to pick: For paraffin-section IHC, choose A04444-3 for its documented EDTA pH 8.0 retrieval and 2 μg/ml staining conditions, or PA2216 for its human liver cancer IHC image and listed 0.5–1 μg/ml range (A04444-3 IHC captions; PA2216 IHC caption and catalog dilution). For IF, choose A04444-3 at its listed 5 μg/ml; ICC validation is unreported (A04444-3 IF caption; catalog applications). For cross-species studies, A04444-3 lists human, mouse and rat reactivity, while its IHC-P dilution specifies human and rat; both antibodies have unreported clone type and section fixative (catalog reactivity, dilution and clone fields; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13228 (SBP1_HUMAN, Methanethiol oxidase).
- Human Protein Atlas. SELENBP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SELENBP1 subcellular location (ICC-IF): Localized to the nucleoli..
- Human Protein Atlas. SELENBP1 antibody validation summary (3 antibodies).
- The function and significance of SELENBP1 downregulation in human bronchial epithelial carcinogenic process. PloS one 2013 — PMC3747066.
- Selenium-binding protein 1 is down-regulated in malignant melanoma. Oncotarget 2018 — PMC5828193.
- Selenium-binding protein 1 in head and neck cancer is low-expression and associates with the prognosis of nasopharyngeal carcinoma. Medicine 2016 — PMC5008557.
- SELENBP1 inhibits progression of colorectal cancer by suppressing epithelial-mesenchymal transition. Open medicine (Warsaw, Poland) 2022 — PMC9438969.
- PubMed PMID:9027582 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SELENBP1 (Methanethiol oxidase). Expected localisation Predominantly cytoplasmic; nuclear in some cases (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04444-3). Validated IHC antibody: A04444-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Endocrine cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human spleen tissue; fixative not specified (datasheet A04444-3); 2. Fixation: Image fixative and duration unreported (datasheet A04444-3); 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 A04444-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04444-3); 8. Primary antibody: Rabbit anti-SELENBP1, 2-5 μg/ml (datasheet A04444-3); 9. Primary incubation: Overnight at 4 °C (datasheet A04444-3). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Lung, colorectal and ovarian tumors may stain less (UniProt); Isoform / epitope: 4 isoforms; epitope impact is undetermined (UniProt); Regulation: Down in lung/colorectal/ovarian cancer (UniProt). Evidence: UniProt Q13228, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoli., Datasheet A04444-3, PMC5828193, PMC5008557. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SELENBP1/immunohistochemistry-ihc-guide.