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SCNN1B / Epithelial sodium channel subunit beta · IHC design guide
Design Immunohistochemistry for SCNN1B
Plan SCNN1B staining in paraffin sections using reported cytoplasmic tissue staining and the expected apical membrane location (HPA tissue IHC; UniProt). Compare appropriate tissue controls and interpret staining cautiously because antibody staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SCNN1B 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 in tissue (HPA tissue IHC); apical membrane expected (UniProt) | |
| Staining pattern | Cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A01670) | |
| Positive control | Stomach+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections (selected-SKU IHC image A01670) | |
| Caveat | Tissue staining has low concordance with RNA (HPA tissue IHC) | |
| Regulation | pro-PCSK9 promotes ENaC degradation (UniProt) | |
| Isoform / epitope | 2 isoforms; check extracellular versus cytoplasmic epitope (UniProt) |
Section 1
Recommended SCNN1B IHC & IF Protocols
The catalog antibody protocol uses heat-mediated Tris-EDTA retrieval at pH 8.0 (datasheet A01670). The published colorectal tissue protocol below provides IHC staining details (PMC11484387).
| Sample | Paraffin-embedded Human colon cancer tissue; fixative not specified (datasheet A01670) |
| Fixation | Image fixative and duration unreported (datasheet A01670); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0 (datasheet A01670); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SCNN1B, 1:100-1:300 (datasheet A01670) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SCNN1B-positive staining in glandular cells of stomach (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA retrieval at pH 8.0 (datasheet A01670); the published protocol does not specify retrieval conditions (PMC11484387).
Section 2
What Is the Expected SCNN1B Staining Pattern?
SCNN1B is an apical cell membrane protein that can also occur on cytoplasmic vesicle membranes (UniProt P51168 localization; topology: 2 transmembrane segments). In paraffin sections, examine epithelial cells, especially kidney tubules and bronchial respiratory epithelium, and stomach glandular cells (HPA: medium, medium and high staining, respectively). HPA describes cytoplasmic staining across several tissues, but rates its tissue IHC profile Uncertain because staining and RNA expression have low consistency (HPA: tissue IHC).
| Apical-edge staining in kidney tubular cells, with or without adjacent cytoplasmic staining. | This fits the expected membrane compartment (UniProt P51168: apical cell membrane) and an observed positive cell population (HPA: medium in kidney tubular cells). Cytoplasmic staining can be compatible with vesicle localization (UniProt P51168: cytoplasmic vesicle membrane), but HPA's uncertain tissue profile means the pattern alone does not verify antibody specificity (HPA: tissue IHC, Uncertain). |
| Predominantly cytoplasmic staining in stomach glandular cells, without a clearly resolved apical rim. | HPA reports high staining in stomach glandular cells and describes cytoplasmic expression in several tissues (HPA: tissue IHC). Interpret this as an observed IHC pattern, while noting that UniProt places SCNN1B at the apical membrane and on cytoplasmic vesicles (UniProt P51168 localization). Cytoplasmic signal alone cannot establish which structure contains the antigen (general IHC interpretation). |
| Strong, isolated nuclear staining or staining concentrated in an unrelated compartment. | That distribution does not match the annotated apical membrane or cytoplasmic vesicle membrane locations (UniProt P51168 localization). Treat it as a possible artefact and compare with tissue morphology and controls before assigning SCNN1B identity (general IHC practice); HPA's Uncertain reliability makes compartment agreement especially useful, though it is not proof (HPA: tissue IHC). |
| Staining in cells reported as negative, such as bone marrow hematopoietic cells or colon endothelial cells. | These populations are reported as not detected in HPA tissue IHC (HPA: bone marrow hematopoietic cells; colon endothelial cells). Unexpected chromogen may reflect cross-reactivity or endogenous detection activity (general IHC practice). Check a no-primary control and the detection workflow; because HPA rates its profile Uncertain, do not treat its negative calls as absolute biological exclusions (HPA: tissue IHC). |
| Uniform haze across epithelium and surrounding tissue, or no signal in a kidney section. | Uniform haze without cell or compartment structure is difficult to assign to SCNN1B (general IHC interpretation). Kidney tubular cells are an observed medium positive population, and kidney protein expression is reported independently by UniProt (HPA: kidney tubular cells; UniProt P51168: kidney protein expression). A blank kidney run warrants a technical check, but cannot by itself prove absence of SCNN1B. |
💡Expected SCNN1B appearanceA persuasive positive shows epithelial staining with an apical component where resolved, such as medium kidney tubular staining, while high stomach glandular staining may appear cytoplasmic; isolated nuclear signal or uniform tissue haze is suspect, and all calls remain provisional because HPA rates tissue IHC Uncertain (UniProt P51168 localization; HPA: kidney, stomach and tissue IHC; general IHC interpretation).
| Membrane topology and epitope position. | SCNN1B has cytoplasmic residues 1–50 and 554–640, an extracellular region at 72–532, and 2 transmembrane segments (UniProt P51168 topology). An antibody's epitope position would affect how staining is interpreted, but no epitope is supplied here; do not infer its accessibility or prescribe antigen retrieval from topology alone. |
| Observed tissue pattern and reliability. | HPA reports high stomach glandular staining and medium staining in kidney tubular and bronchial respiratory epithelial cells, yet labels the overall tissue IHC profile Uncertain because of low staining–RNA consistency (HPA: tissue IHC). Use these populations as comparison tissues, with morphology and controls, rather than as definitive specificity standards (general IHC practice). |
| Cytoplasmic signal alongside expected membrane localization. | UniProt lists both apical cell membrane and cytoplasmic vesicle membrane locations (UniProt P51168 localization); HPA describes cytoplasmic expression in several tissues (HPA: tissue IHC). Record membrane and cytoplasmic compartments separately when scoring (general IHC practice). The supplied evidence does not establish that every cytoplasmic deposit represents vesicular SCNN1B. |
| Isoforms and antibody validation. | UniProt lists 2 isoforms (UniProt P51168 isoforms). The supplied records do not identify the catalog antibody's epitope or isoform coverage, so staining cannot be assigned to a particular isoform. The listed HPA antibody, HPA015612, has an Uncertain IHC status (HPA: antibody validation); compare results with independent evidence before making a strong specificity claim (general IHC practice). |
| IF/ICC comparison. | HPA summarizes the subcellular location as membrane but provides no main location or cell lines with ICC-IF images (HPA: subcellular record). This offers limited support for a membrane expectation; it does not validate an IF/ICC image pattern. Address IF/ICC setup and interpretation in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubules show no staining. | A weak run is possible because HPA reports medium staining in tubular cells; tissue variation also remains possible under HPA's Uncertain profile (HPA: kidney and tissue IHC). | Confirm that tubules are present, inspect the run's positive and negative controls, and review antibody dilution, retrieval and chromogen steps as general IHC checks (general IHC practice). Do not infer a SCNN1B-specific retrieval requirement from these sources. |
| Stomach glandular cells are blank in an intended positive comparison. | HPA reports high staining in this cell population, so a blank result raises a run or specimen question; HPA's uncertain reliability prevents treating one section as a guaranteed positive (HPA: stomach; tissue IHC). | Verify glandular morphology and control performance, then review primary incubation and detection reagents (general IHC practice). Compare another documented positive population, such as kidney tubules, if available (HPA: kidney tubular cells, medium). |
| Signal appears chiefly in nuclei. | A nuclear-only pattern conflicts with the annotated apical and vesicle membrane locations (UniProt P51168 localization); nonspecific binding or chromogen artefact is possible (general IHC practice). | Compare the nuclear signal with a no-primary control, check counterstain and precipitate, and score membrane and cytoplasmic staining separately (general IHC practice). Do not call nuclear staining a confirmed SCNN1B pattern from the supplied evidence. |
| Bone marrow hematopoietic cells stain strongly. | HPA reports these cells as not detected (HPA: bone marrow hematopoietic cells). Cross-reactivity or endogenous peroxidase activity could account for unexpected chromogen in an IHC run (general IHC practice). | Inspect a no-primary control and the peroxidase-blocking step, then compare the distribution with a reported positive epithelial population (general IHC practice; HPA: kidney tubular cells, medium). Treat the HPA negative call as provisional because tissue reliability is Uncertain (HPA: tissue IHC). |
| Diffuse background obscures cell borders. | Indistinct staining prevents reliable compartment scoring (general IHC interpretation); the supplied records do not identify a SCNN1B-specific cause. | Review blocking, washing, primary antibody dilution and chromogen development against the run controls (general IHC practice). Reassess apical and cytoplasmic distribution only after tissue structures are readable (UniProt P51168 localization; general IHC practice). |
| Can IF/ICC confirm an apical SCNN1B pattern? | HPA provides a membrane summary but no ICC-IF image-bearing cell lines or specified main location (HPA: subcellular record). | Use the separate IF/ICC guide for that application. Interpret any IF/ICC image with compartment markers and controls as general imaging practice; the supplied HPA entry alone cannot confirm an apical IF/ICC pattern. |
Experiment evidence
IHC & IF images showing SCNN1B staining
A01670 · IHCImmunohistochemical analysis of paraffin-embedded Human colon cancer. Antibody was diluted at 1:100 (4° overnight). High-pressure and temper
A01670 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using Nonvoltage-gated Sodium Channel 1 Antibody. The picture on the
A01670 · IFImmunofluorescence analysis of HUVEC cells, using Nonvoltage-gated Sodium Channel 1 Antibody. The picture on the right is blocked with the s
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168447-SCNN1B/tissue/cerebral+cortex#imid_4071060
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168447-SCNN1B/tissue/colon#imid_4071002
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168447-SCNN1B/tissue/liver#imid_4070987
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000168447-SCNN1B/tissue/kidney#imid_4070988
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SCNN1B IHC & IF
🧪Run stomach first and look for staining in glandular cells (HPA: High in stomach glandular cells). Use rectum glandular cells as the negative tissue (HPA: Not detected in rectum glandular cells); on the stomach slide, adjacent nonglandular cells should show only background staining and serve as an internal background reference (UniProt P51168: epithelial channel).
Positive control tissue: Stomach (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for SCNN1B; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, immunoglobulin class and clonality; and a peptide-block control using the immunogen peptide (selected-SKU tissue-IHC caption: peptide pre-absorption control). Check for endogenous peroxidase staining in the stomach section before interpreting chromogenic signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected-SKU paraffin-section caption is unreported (selected-SKU tissue-IHC caption). That caption uses Tris-EDTA at pH 8.0 for heat retrieval in colon cancer, but does not establish whether retrieval is required for stomach or whether frozen sections or IF are easier (selected-SKU tissue-IHC caption). In stomach, evaluate glandular staining against the no-primary control to distinguish specific signal from endogenous peroxidase background (HPA: High in stomach glandular cells; standard IHC practice).
HPA tissue IHC evidence for SCNN1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Colon | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Rectum | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SCNN1B IHC Tips
Use compartment aware controls and matched processing to troubleshoot SCNN1B staining in paraffin sections (UniProt P51168 topology; datasheet A01670).
How should I adjust retrieval when SCNN1B staining is weak?
Start with heat mediated Tris-EDTA at pH 8.0 for SCNN1B paraffin section IHC (datasheet A01670). The selected image used high pressure and temperature retrieval, but gives no heating duration, so record the instrument, temperature, and time used in your laboratory (caption A01670). If signal is weak, compare a modestly longer heating interval with the original condition on adjacent sections, while keeping antibody dilution and detection constant (standard IHC practice). Check section integrity and background after each change; excessive retrieval can damage morphology and make apparent membrane staining difficult to judge (standard IHC practice).
Could fixation explain variable SCNN1B staining between paraffin blocks?
Target specific sensitivity of SCNN1B to fixation is unknown from the supplied evidence; the selected paraffin section caption does not report a fixative (caption A01670). Record fixative, fixation duration, tissue thickness, and processing history for each block before attributing different staining to biology (standard IHC practice). Compare blocks with similar processing using the same retrieval, antibody concentration, and development time, and include a reference section in each run (standard IHC practice). If heavily processed material stains weakly, assess morphology and an independent tissue control before changing retrieval, since processing differences can alter antigen accessibility (standard IHC practice).
Which SCNN1B staining pattern should I accept as plausible?
Prioritize a polarized apical membrane pattern in epithelial cells, while allowing intracellular vesicular signal as a possibility (UniProt P51168 subcellular location). SCNN1B has 2 transmembrane segments, at residues 51–71 and 533–553, with a large extracellular region between them (UniProt P51168 topology). HPA reports cytoplasmic staining across several tissues, but rates tissue IHC reliability uncertain because staining and RNA expression show low consistency (HPA tissue IHC). Evaluate the membrane boundary against tissue architecture and counterstain, then compare suspicious diffuse signal with a negative control processed in the same run (standard IHC practice).
Can epitope placement or isoforms explain discordant SCNN1B IHC?
The record lists 2 SCNN1B isoforms, but supplies no antibody epitope or isoform coverage; do not assume this antibody detects both (UniProt P51168 isoforms; caption A01670). Map the immunogen sequence, if available, against the extracellular 72–532 region and cytoplasmic 1–50 and 554–640 regions before interpreting staining differences (UniProt P51168 topology). The record also identifies glycosylation at residue 260 and phosphoserines at 633 and 635, without establishing an effect on this antibody (UniProt P51168 modifications). Compare an independently mapped antibody or orthogonal expression evidence when epitope accessibility remains uncertain (standard IHC validation practice).
How can IF help assess ambiguous SCNN1B IHC localisation?
Use IF on matched material to assess whether SCNN1B signal follows epithelial cell boundaries or an intracellular pattern suggested by the IHC section (UniProt P51168 subcellular location; standard IF practice). Multiplex with a validated epithelial marker appropriate to the sampled tissue and choose spectrally separated fluorophores after checking tissue autofluorescence in an unstained section (standard IF practice). Set permeabilisation according to the antibody’s mapped epitope: a cytoplasmic epitope requires membrane access, whereas an extracellular epitope may be assessed with minimal permeabilisation (UniProt P51168 topology; standard IF practice). Because no SCNN1B ICC/IF images or epitope mapping are supplied here, validate this IF readout independently before using it to resolve IHC discrepancies (HPA subcellular record; caption A01670).
What should I check when chromogenic SCNN1B staining is diffuse?
Inspect a no primary control for detection system background and an adjacent stained section for preserved morphology before increasing antibody concentration (standard IHC practice). Apply a peroxidase block when using an HRP and DAB workflow, and check whether endogenous enzyme activity persists in the control (standard chromogenic IHC practice). The selected image used antibody at 1:100, incubated at 4°C overnight, and included immunogen peptide preabsorption as a negative control (caption A01670). Peptide competition can support epitope dependent binding, but diffuse staining still needs assessment against tissue architecture and independent controls (caption A01670; standard IHC validation practice).
How should I score SCNN1B staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible epithelial compartment and an apical membrane scoring rule before reviewing study groups (UniProt P51168 subcellular location; standard IHC scoring practice). Record the percentage of positive eligible cells and intensity, or calculate an H-score from intensity categories and their cell percentages (standard IHC scoring practice). Normalize counts to the number of eligible epithelial cells; if measuring stained area or density per mm², normalize to viable epithelial area rather than total section area (standard image analysis practice). Keep retrieval, antibody incubation, chromogen development, imaging, and thresholds consistent, and report cytoplasmic and membrane signals separately because HPA tissue IHC is rated uncertain (caption A01670; HPA tissue IHC; standard IHC practice).
How do I distinguish convincing SCNN1B signal from artefact?
Favor reproducible staining in the expected epithelial compartment, especially along the apical surface, and assess cytoplasmic staining cautiously because HPA reports it with uncertain reliability (UniProt P51168 subcellular location; HPA tissue IHC). Kidney tubule cells and bronchial respiratory epithelial cells have reported medium staining, while colon endothelial cells are reported as not detected (HPA tissue IHC). Compare suspected signal with a no primary control and inspect section edges, necrotic regions, and endogenous enzyme activity for artefactual chromogen deposition (standard IHC practice). The peptide preabsorption control in the selected image supports antibody dependent staining there, but does not establish specificity in every tissue or compartment (caption A01670; standard IHC validation practice).
Boster reagents
Best SCNN1B / Epithelial sodium channel subunit beta IHC Antibodies
A01670 has IHC images of paraffin-embedded human colon cancer and brain tissue, plus an IF image of HUVEC cells (catalog image captions). Listed reactivity is human, mouse, and rat (catalog).
Real IHC data
Anti-ENaC beta SCNN1B Antibody
Cat # A01670
A01670 is listed for IHC and IF, with IHC images of paraffin-embedded human colon cancer and brain tissue (catalog applications; IHC image captions). Its IF image shows HUVEC cells; the catalog lists human, mouse, and rat reactivity (IF image caption; catalog reactivity).
Which to pick: For tissue IHC, choose A01670 based on its paraffin-section images; the fixative is unreported (IHC image captions). For IF/ICC work, A01670 has an IF image in HUVEC cells, but ICC validation and clonality are unreported (IF image caption; catalog). For cross-species work, A01670 lists human, mouse, and rat reactivity, while the supplied IHC and IF images show human samples only (catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51168 (SCNNB_HUMAN, Epithelial sodium channel subunit beta).
- Human Protein Atlas. SCNN1B tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. SCNN1B subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SCNN1B antibody validation summary (1 antibodies).
- Hypermethylation of the sodium channel beta subunit gene promoter is associated with colorectal cancer. Hereditas 2024 — PMC11484387.
- Genes associated with inflammation for prognosis prediction for clear cell renal cell carcinoma: a multi-database analysis. Translational cancer research 2023 — PMC10643973.
- The sodium channel subunit SCNN1B suppresses colorectal cancer via suppression of active c-Raf and MAPK signaling cascade. Oncogene 2023 — PMC9937924.
- Correlating obstructive sleep apnea and lung adenocarcinoma using hub gene signatures and molecular mechanisms. Discover oncology 2026 — PMC13338089.
- PubMed PMID:7490094 — UniProt-cited evidence.
- PubMed PMID:7762608 — UniProt-cited evidence.
- PubMed PMID:9813171 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SCNN1B (Epithelial sodium channel subunit beta). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); apical membrane expected (UniProt). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A01670). Validated IHC antibody: A01670 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Stomach (Glandular cells); negative Bone marrow. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human colon cancer tissue; fixative not specified (datasheet A01670); 2. Fixation: Image fixative and duration unreported (datasheet A01670); 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: Tris-EDTA pH 8.0 (datasheet A01670); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SCNN1B, 1:100-1:300 (datasheet A01670); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across compared paraffin sections (selected-SKU IHC image A01670); Caveat: Tissue staining has low concordance with RNA (HPA tissue IHC); Isoform / epitope: 2 isoforms; check extracellular versus cytoplasmic epitope (UniProt); Regulation: pro-PCSK9 promotes ENaC degradation (UniProt). Evidence: UniProt P51168, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A01670, PMC11484387. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SCNN1B/immunohistochemistry-ihc-guide.