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SBNO1 / Protein strawberry notch homolog 1 · IHC design guide
Design Immunohistochemistry for SBNO1
Plan SBNO1 IHC-P around nuclear staining in most tissues (HPA tissue IHC). Use appendix glandular cells, which show high staining (HPA tissue IHC), as a positive tissue reference when interpreting chromogenic signal.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SBNO1 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 | Nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A13130) | |
| Positive control | Adrenal gland+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 | Keep paraffin-section 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 A13130) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | RNA is tissue enhanced in testis (HPA tissue RNA) | |
| Isoform / epitope | Four isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended SBNO1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published human CCA tissue protocol (PMC13067845).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A13130) |
| Fixation | Image fixative and duration unreported (datasheet A13130); 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 A13130); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A13130) |
| Primary antibody | Rabbit anti-SBNO1, 2-5 μg/ml (datasheet A13130) |
| Primary incubation | Overnight at 4 °C (datasheet A13130) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A13130) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SBNO1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: 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 A13130); the published CCA method specifies heat retrieval without buffer details (PMC13067845).
Section 2
What Is the Expected SBNO1 Staining Pattern?
SBNO1 should appear predominantly in nuclei across most tissues (HPA tissue IHC: nuclear expression in most tissues; UniProt A3KN83: nucleus). High staining is reported in several glandular, respiratory epithelial, hematopoietic, and endothelial cell populations (HPA tissue IHC: High in the listed populations). UniProt annotates no transmembrane segment (UniProt A3KN83 topology). Treat the tissue pattern as a guide, with medium consistency between staining and RNA data and external verification pending (HPA tissue IHC: Approved).
| Distinct nuclear chromogen in adrenal or breast glandular cells, bronchial respiratory epithelial cells, or bone marrow hematopoietic cells. | This fits the reported compartment and high-staining populations (HPA tissue IHC: nuclear expression in most tissues; High in those cells). Compare cells within the section before scoring: the HPA level describes those populations, not every cell in the tissue (HPA tissue IHC). |
| Chromogen is mainly cytoplasmic or outlines cell membranes while nuclei remain pale. | That distribution conflicts with the reported nuclear pattern (HPA tissue IHC: nuclear expression; UniProt A3KN83: nucleus, no transmembrane segment). Review counterstain alignment and detection controls before assigning a cellular compartment (general IHC practice). |
| The strongest staining is in a cell population other than the documented high-staining population within the same tissue. | Check cell identity and possible cross-reactivity or endogenous detection activity (general IHC practice). This is a discrepancy, not proof of a false positive: HPA reports nuclear expression in most tissues and lists selected high-staining populations (HPA tissue IHC). |
| Weak, diffuse chromogen covers tissue structures and obscures nuclear boundaries. | Interpretation is unreliable when background prevents nuclear scoring (general IHC practice). Compare a no-primary control and reassess blocking, antibody concentration, washing, and chromogen development (general IHC practice). |
| No nuclear signal appears in a section expected to contain a listed high-staining population. | First confirm that the relevant cells are present; examples include appendix glandular cells or colon endothelial cells (HPA tissue IHC: High). If present, investigate assay performance with a suitable positive section and detection controls (general IHC practice). |
💡Expected SBNO1 appearanceCall positive a clearly bounded nuclear signal in an HPA-listed high-staining cell population, such as duodenal glandular cells; HPA reports that population as High, while dominant membrane staining conflicts with the nuclear annotation (HPA tissue IHC; UniProt A3KN83 topology).
| Compartment and topology | Nuclear staining is the primary localisation criterion (UniProt A3KN83: nucleus; HPA tissue IHC: nuclear expression in most tissues). No transmembrane segment is annotated, so a dominant membrane outline needs investigation (UniProt A3KN83 topology). |
| Choice of comparison tissue | Adrenal, appendix, breast, cervix, and duodenal glandular cells; bronchial respiratory epithelial cells; bone marrow hematopoietic cells; and colon endothelial cells are listed as High (HPA tissue IHC). Glial cells in hippocampus and caudate are listed as Low, making them poor choices as sole positive controls (HPA tissue IHC). |
| Evidence strength | The tissue profile is Approved, with medium consistency between antibody staining and RNA expression and external verification pending (HPA tissue IHC). HPA042388 is IHC Approved; the supplied antibody record does not report Enhanced IHC validation (HPA antibodies). Interpret unexpected patterns with appropriate controls (general IHC practice). |
| Isoforms and antibody coverage | Four SBNO1 isoforms are listed (UniProt A3KN83: isoforms 1–4). The supplied record does not map the IHC antibody epitope or establish detection of each isoform (HPA antibodies; UniProt A3KN83). Do not attribute staining differences to a particular isoform from these data alone. |
| IF/ICC question: where should signal appear? | In ICC-IF, nucleoplasmic signal is the approved HPA localisation; image lines listed are A-431, A-549, U2OS, and NIH 3T3 (HPA subcellular; HPA antibodies: HPA042388 ICC Approved). This supports a localisation check, not an IF protocol or a tissue IHC intensity prediction. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed high-staining population shows no nuclear chromogen. | The relevant cells may be absent from the section, or an assay step may have failed (general IHC practice); the population is reported High, not guaranteed positive in every specimen (HPA tissue IHC). | Confirm cell identity and preservation, then run a suitable positive section and detection controls; review the antibody's documented IHC-P conditions (general IHC practice). |
| Signal appears mainly in cytoplasm or along cell borders. | The observed compartment disagrees with nuclear SBNO1 localisation (UniProt A3KN83: nucleus; HPA tissue IHC: nuclear expression). Background or misassigned cell boundaries may complicate scoring (general IHC practice). | Check the nuclear counterstain and no-primary control, then score only signal confidently assigned to nuclei (general IHC practice). |
| Brown deposit is diffuse across the section. | Excess background, including insufficient blocking or washing, can obscure cellular localisation (general IHC practice). The supplied sources do not establish SBNO1-specific background behaviour (HPA tissue IHC; UniProt A3KN83). | Compare a no-primary control; review blocking, wash steps, antibody concentration, and development time under the assay's standard workflow (general IHC practice). |
| Unexpected cells stain strongly within a tissue. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). The HPA list names selected High populations and does not define every other population as negative (HPA tissue IHC). | Confirm cell identity and nuclear localisation; use no-primary and detection-system controls before calling the staining target specific (general IHC practice). |
| A hippocampal or caudate glial section gives little signal. | Glial cells in those regions are listed as Low (HPA tissue IHC); weak staining there alone does not demonstrate assay failure. | Assess a section containing an HPA-listed High population and verify the expected nuclear pattern before adjusting the assay (HPA tissue IHC; general IHC practice). |
| An IF/ICC image shows a bright nonnuclear pattern. | That pattern conflicts with the approved nucleoplasmic ICC-IF localisation (HPA subcellular). Autofluorescence or nonspecific signal can also affect IF interpretation (general IF practice). | Check nuclear counterstain registration and appropriate fluorescence controls, then compare localisation with the HPA nucleoplasmic reference (general IF practice; HPA subcellular). |
Experiment evidence
IHC & IF images showing SBNO1 staining
A13130 · IHCIHC analysis of SBNO1 using anti-SBNO1 antibody (A13130). SBNO1 was detected in a paraffin-embedded section of human appendiceal adenocarcin
A13130 · IHCIHC analysis of SBNO1 using anti-SBNO1 antibody (A13130). SBNO1 was detected in a paraffin-embedded section of human breast cancer tissue. H
A13130 · IFIF analysis of SBNO1 using anti-SBNO1 antibody (A13130). SBNO1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen r
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000139697-SBNO1/tissue/cerebral+cortex#imid_11153328
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000139697-SBNO1/tissue/colon#imid_11210397
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000139697-SBNO1/tissue/liver#imid_11210473
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000139697-SBNO1/tissue/kidney#imid_11210414
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042388; green = SBNO1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042388; green = SBNO1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SBNO1 IHC & IF
🧪Run appendix first and assess its glandular cells for SBNO1 staining (HPA: High in appendix glandular cells). HPA detects SBNO1 in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and treat cells without nuclear signal on the positive slide as background comparators, not confirmed SBNO1-negative cells (HPA: no negative tissue rows; UniProt A3KN83: nucleus).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: SBNO1 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 SBNO1 in A-431, A-549, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a host- and clonality-matched rabbit IgG isotype control, and a knockout specimen or validated peptide-block control where available (selected A13130 caption: rabbit primary antibody; standard IHC control practice). Quench endogenous peroxidase and assess endogenous biotin background in appendix because the reported detection uses a biotinylated secondary, streptavidin complex and DAB (selected A13130 caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A13130 paraffin-section caption does not state a fixative (selected A13130 caption: fixative unreported). The caption reports heat retrieval in EDTA at pH 8.0, but one reported condition does not establish that retrieval is required (selected A13130 caption). HPA ICC-IF images support assessing nucleoplasmic signal, but the supplied evidence does not establish that frozen sections or IF are easier; for the appendix IHC protocol, check biotin-associated background from the reported detection system (HPA: nucleoplasm approved; selected A13130 caption: streptavidin-biotin detection).
HPA tissue IHC evidence for SBNO1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SBNO1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SBNO1 IHC Tips
Troubleshoot SBNO1 staining by checking nuclear localisation, the validated retrieval conditions, and matched controls before comparing tissue scores.
What retrieval should I use when nuclear SBNO1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A13130). The selected antibody stained a paraffin-embedded human appendiceal adenocarcinoma section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A13130). If nuclei remain weak, vary heating duration within the retrieval equipment’s recommended range and compare serial sections processed together (standard IHC practice). Check tissue preservation and a positive control before raising primary concentration, since uneven heating or tissue loss can resemble poor epitope exposure (standard IHC practice). Record retrieval conditions alongside nuclear signal and tissue damage for each trial (standard IHC practice).
Could fixation explain variable SBNO1 staining between paraffin sections?
Target-specific SBNO1 sensitivity to fixative choice or fixation duration is unknown from the supplied evidence; the selected tissue caption reports paraffin embedding but no fixative (datasheet A13130). Compare sections with documented fixation histories under the same EDTA pH 8.0 retrieval and detection conditions (datasheet A13130; standard IHC practice). Excessive or inconsistent fixation can alter antigen accessibility in general, so treat any resulting signal change as an experimental observation rather than an established SBNO1 effect (standard IHC practice). Include similarly processed control tissue in each run and assess nuclear signal together with morphology (standard IHC practice). Do not infer fixation tolerance from tissue expression patterns or protein modifications (HPA tissue IHC; UniProt A3KN83).
How should I troubleshoot cytoplasmic staining when SBNO1 is expected in nuclei?
Evaluate SBNO1 primarily in nuclei: UniProt assigns it to the nucleus, and HPA reports approved nucleoplasmic localisation (UniProt A3KN83; HPA subcellular). HPA also describes nuclear expression in most tissues, although its tissue staining has medium consistency with RNA expression and awaits external verification (HPA tissue IHC). Compare nuclear and cytoplasmic signal in intact cells at the same illumination and staining conditions, rather than scoring total brown area (standard IHC practice). If cytoplasmic colour dominates, inspect no-primary and detection controls for nonspecific deposition, then titrate the primary antibody (standard IHC practice). Do not treat isolated cytoplasmic staining as confirmed SBNO1 localisation without independent validation (UniProt A3KN83; HPA subcellular).
Can isoforms or modifications account for inconsistent SBNO1 nuclear staining?
SBNO1 has 4 annotated isoforms and multiple modified residues, including phosphorylation and acetylation sites (UniProt A3KN83). The supplied caption does not identify the A13130 epitope or establish which isoforms it detects, so staining differences cannot be assigned to a particular variant (datasheet A13130; UniProt A3KN83). Request or check the immunogen sequence and map it against isoforms before interpreting a negative section as isoform absence (standard antibody validation practice). Compare matched sections under the stated EDTA pH 8.0 retrieval while holding detection conditions constant (datasheet A13130; standard IHC practice). If variant specificity matters, corroborate with an independently characterised epitope or orthogonal assay (standard antibody validation practice).
How can IF help check an ambiguous chromogenic SBNO1 result?
Use IF as an independent localisation check: HPA reports SBNO1 in the nucleoplasm, while the selected A13130 tissue caption documents chromogenic IHC rather than IF validation (HPA subcellular; datasheet A13130). Multiplex SBNO1 with a validated marker for the expected cell population, such as glandular cells when examining appendix, and include a nuclear counterstain (HPA tissue IHC; standard IF practice). Choose a spectrally separated fluorophore, preferably in a lower-autofluorescence channel, after inspecting unstained tissue and single-label controls (standard IF practice). Because the expected signal is nuclear and SBNO1 has no annotated transmembrane segment, optimise mild permeabilisation for nuclear antibody access after fixation (UniProt A3KN83; standard IF practice). Compare nuclear overlap with the chromogenic pattern without assuming A13130 has been validated for IF (datasheet A13130).
What causes diffuse brown background in an SBNO1 IHC section?
First distinguish diffuse colour from the predominantly nuclear pattern expected for SBNO1 (UniProt A3KN83; HPA tissue IHC). The selected caption uses 10% goat serum blocking, 2 μg/ml rabbit primary antibody, a biotinylated secondary, an avidin-biotin complex, and DAB (datasheet A13130). Run no-primary and secondary-only controls, block endogenous peroxidase, and assess endogenous biotin when using this detection system (standard IHC practice). If background persists, titrate primary antibody, improve washes, and check whether DAB development is excessive while keeping control sections alongside the test (standard IHC practice). Score nuclei only after background is acceptably low and morphology remains intact (standard IHC practice).
How should I score SBNO1 across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear scoring threshold before comparing slides, because the expected SBNO1 pattern is nuclear (UniProt A3KN83; HPA tissue IHC). Within each compartment, report the percentage of positive nuclei and an H-score based on intensity; a density per mm² can supplement these measures when cell abundance varies (standard IHC scoring practice). Normalise positive counts to evaluable nuclei of the same cell type, and exclude necrotic or poorly preserved areas (standard IHC scoring practice). Keep retrieval, staining batch, counterstain, imaging, and threshold settings consistent across compared sections (standard IHC practice). Report tissue identity and cell type separately, since HPA records different high-staining populations across tissues (HPA tissue IHC).
Which SBNO1 staining patterns deserve validation before biological interpretation?
A convincing result shows reproducible nuclear staining in morphologically intact cells, consistent with UniProt’s nuclear assignment and HPA’s approved nucleoplasmic localisation (UniProt A3KN83; HPA subcellular). Consider cell identity: HPA reports high staining in appendix glandular cells but low staining in hippocampal and caudate glial cells (HPA tissue IHC). Predominantly cytoplasmic colour, section-edge enhancement, necrotic deposits, or signal in no-primary controls calls for investigation of background and detection artefacts (standard IHC practice). Check endogenous peroxidase and, with the caption’s avidin-biotin detection, possible endogenous biotin before attributing brown signal to SBNO1 (datasheet A13130; standard IHC practice). Interpret expression cautiously because the HPA tissue profile has medium RNA-staining consistency and awaits external verification (HPA tissue IHC).
Boster reagents
Best SBNO1 / Protein strawberry notch homolog 1 IHC Antibodies
The catalog lists one anti-SBNO1 antibody with paraffin-section IHC images from human tumors and an IF image from A549 cells (catalog: A13130 image captions).
Real IHC data
Anti-SBNO1 Antibody ®
Cat # A13130
A13130 has IHC images from human appendiceal adenocarcinoma, breast cancer, esophageal squamous carcinoma, and gall bladder adenosquamous carcinoma paraffin sections (catalog: A13130 IHC captions). A13130 also has an IF image from A549 cells and lists Human, Mouse, and Rat reactivity (catalog: A13130 IF caption and reactivity).
Which to pick: For tissue IHC, choose A13130: its paraffin-section captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (catalog: A13130 IHC captions). For IF/ICC, A13130 has an A549 IF image using 5 μg/ml primary antibody (catalog: A13130 IF caption). A13130 is the only listed cross-species candidate, with Human, Mouse, and Rat reactivity; its host is Rabbit, while clonality and mouse or rat IHC image evidence are unreported (catalog: A13130 reactivity, host, clone, and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry A3KN83 (SBNO1_HUMAN, Protein strawberry notch homolog 1).
- Human Protein Atlas. SBNO1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SBNO1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. SBNO1 antibody validation summary (1 antibodies).
- Strawberry notch 1 safeguards neuronal genome via regulation of Yeats4 expression. Cell death discovery 2025 — PMC12289961.
- Strawberry Notch 1 Acts as a Transcriptional Regulator Driving Oncogenic Programs in Liver Carcinogenesis. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13067845.
- Neuronal Populations Involved in Motor Function Show Prominent Expression of Sbno1 During Postnatal Brain Development. Journal of developmental biology 2025 — PMC11843823.
- Sbno1 mediates cell-cell communication between neural stem cells and microglia through small extracellular vesicles. Cell & bioscience 2024 — PMC11441009.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16533400 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SBNO1 (Protein strawberry notch homolog 1). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A13130). Validated IHC antibody: A13130 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 appendiceal adenocarcinoma tissue; fixative not specified (datasheet A13130); 2. Fixation: Image fixative and duration unreported (datasheet A13130); 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 A13130); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A13130); 8. Primary antibody: Rabbit anti-SBNO1, 2-5 μg/ml (datasheet A13130); 9. Primary incubation: Overnight at 4 °C (datasheet A13130). Watch for: Fixation: Keep paraffin-section 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 A13130); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: Four isoforms; epitope coverage is unreported (UniProt); Regulation: RNA is tissue enhanced in testis (HPA tissue RNA). Evidence: UniProt A3KN83, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A13130, PMC13067845. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SBNO1/immunohistochemistry-ihc-guide.