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SCN4B / Sodium channel regulatory subunit beta-4 · IHC design guide
Design Immunohistochemistry for SCN4B
Plan chromogenic SCN4B IHC in paraffin sections using the variable cytoplasmic tissue pattern as a reference (HPA tissue IHC). Start with the catalog antibody's 0.5–1 μg/mL IHC-P range (datasheet A06453-1), and assess membrane staining in light of the annotated cell-membrane location (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SCN4B Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasm observed (HPA tissue IHC); membrane annotated (UniProt) | |
| Staining pattern | Variable cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A06453-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06453-1) | |
| Caveat | IHC staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Higher expression in dorsal root ganglia (UniProt) | |
| Isoform / epitope | 3 isoforms; map the epitope to the extracellular or cytoplasmic region (UniProt) |
Section 1
Recommended SCN4B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published SCN4B IHC protocol covering human tissue arrays and mouse tumors (PMC5150224 Methods).
| Sample | Paraffin-embedded human renal cancer tissue; fixative not specified (datasheet A06453-1) |
| Fixation | Image fixative and duration unreported (datasheet A06453-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A06453-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06453-1) |
| Primary antibody | Rabbit anti-SCN4B, 0.5-1μg/ml (datasheet A06453-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06453-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06453-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SCN4B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in all tissues at variable levels. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 heat retrieval (datasheet A06453-1); the published human tissue array protocol used Tris/EDTA pH 9.0 (PMC5150224 Methods).
Section 2
What Is the Expected SCN4B Staining Pattern?
SCN4B is a cell-membrane protein with an extracellular region at residues 31–162 and one transmembrane segment at 163–183 (UniProt Q8IWT1 topology). In tissue IHC, HPA reports variable cytoplasmic staining, including medium staining in cerebral cortical neuronal cells (HPA: tissue IHC). Treat compartment calls cautiously: HPA rates its IHC evidence Approved but reports low consistency between antibody staining and RNA expression (HPA: reliability).
| Medium, cell-associated staining in cerebral cortical neuronal cells (HPA: cerebral cortex, Medium). | This matches an observed HPA IHC pattern. Membrane-associated staining is biologically plausible (UniProt Q8IWT1: cell membrane), but HPA describes its tissue IHC pattern as cytoplasmic; membrane enrichment is therefore not a required pass criterion (HPA: tissue IHC). |
| Predominantly nuclear staining, or staining confined to an unrelated subcellular compartment. | That compartment conflicts with SCN4B's annotated cell-membrane location (UniProt Q8IWT1: subcellular location). Check counterstain alignment and detection controls before treating it as specific; misplaced chromogen or nonspecific signal can mislead compartment scoring (general IHC practice). |
| Strong staining in adipocytes or bone-marrow hematopoietic cells. | HPA reports SCN4B as not detected in those cell populations (HPA: adipose tissue; bone marrow). Unexpected signal raises a specificity or endogenous-detection concern; compare a no-primary control and the same staining run's positive tissue before assigning a biological interpretation (general IHC practice). |
| Diffuse color across tissue and empty areas, with little distinction between cells. | A field-wide deposit is difficult to reconcile with HPA's cell-specific observations (HPA: tissue IHC). Review the no-primary control, blocking, washes and chromogen development for background (general IHC practice); HPA's reported cytoplasmic staining alone does not establish an artefact (HPA: tissue IHC). |
| No discernible staining in cerebral cortical neuronal cells. | That misses one HPA-observed medium-staining population (HPA: cerebral cortex, Medium). First verify tissue identity and a working detection control; then review the catalog antibody's IHC-P instructions and the run conditions (general IHC practice). One negative section cannot establish absence of SCN4B. |
💡Expected SCN4B appearanceA defensible positive is medium, cell-associated staining in cerebral cortical neuronal cells, with cytoplasmic appearance acceptable in IHC (HPA: cerebral cortex; tissue IHC); strong adipocyte staining is a specificity flag (HPA: adipocytes, Not detected).
| Compartment evidence | UniProt places SCN4B at the cell membrane, whereas HPA describes variable cytoplasmic tissue IHC staining (UniProt Q8IWT1: subcellular location; HPA: tissue IHC). Record the observed compartment without forcing agreement between these sources. |
| Choice of comparison cells | Cerebral cortical neuronal cells and cerebellar granular-layer cells have medium HPA IHC staining; adipocytes and bone-marrow hematopoietic cells are not detected (HPA: tissue IHC). Compare named cell populations rather than treating an entire section as uniformly positive or negative. |
| Antibody evidence | HPA017293 is IHC Approved, while HPA reports low consistency between antibody staining and RNA expression (HPA: antibody record; reliability). Interpret unexpected cell types conservatively; the supplied record does not establish an IHC Enhanced validation claim. |
| Antigen features | The mature chain begins at residue 31; three glycosylation sites lie in the extracellular region, and UniProt lists three isoforms (UniProt Q8IWT1: processing, topology, glycosylation, isoforms). Without an antibody epitope map, these facts cannot predict retrieval needs or isoform-specific staining. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in an expected positive cell population. | Cerebral cortical neuronal cells show medium staining in HPA, so a blank run needs investigation (HPA: cerebral cortex). | Confirm the cell population and positive detection control; check the catalog antibody's IHC-P instructions, reagent preparation and development time (general IHC practice). |
| Broad, weak haze obscures cell boundaries. | Background from detection or incomplete washing may obscure a cell-associated pattern (general IHC practice). | Compare the no-primary control, then review blocking, washes and chromogen development (general IHC practice); retain HPA's cytoplasmic tissue pattern as a possible specific appearance (HPA: tissue IHC). |
| Strong color appears in adipocytes. | HPA lists adipocytes as not detected; strong signal there is unexpected (HPA: adipose tissue). | Compare a no-primary control and a positive tissue from the same run; assess endogenous detection activity and nonspecific antibody staining before scoring adipocytes as positive (general IHC practice). |
| The apparent signal is mainly nuclear. | Nuclear restriction conflicts with the annotated cell-membrane location (UniProt Q8IWT1: subcellular location). | Check nuclear counterstain alignment, focus and detection controls, then rescore only clearly localized signal (general IHC practice). |
| Cytoplasmic staining appears without a clear membrane outline. | HPA reports cytoplasmic tissue IHC despite UniProt's membrane annotation (HPA: tissue IHC; UniProt Q8IWT1: subcellular location). | Report the observed compartment and cell type; use negative cells and controls to judge specificity. Do not reject the slide solely because membrane enrichment is unresolved (HPA: tissue IHC; general IHC practice). |
| Can an IF/ICC result be judged by this IHC pattern? | HPA summarizes SCN4B as membrane-associated in its subcellular record but supplies no ICC-IF image-bearing cell lines here (HPA: subcellular record). | Treat membrane labeling as a localization expectation (UniProt Q8IWT1: subcellular location; HPA: subcellular summary). Use the separate IF/ICC guide for assay design; this IHC tissue record does not validate an IF staining pattern. |
Experiment evidence
IHC & IF images showing SCN4B staining
A06453-1 · IHCIHC analysis of SCN4B using anti-SCN4B antibody (A06453-1). SCN4B was detected in paraffin-embedded section of human renal cancer tissue. He
A06453-1 · IHCIHC analysis of SCN4B using anti-SCN4B antibody (A06453-1). SCN4B was detected in paraffin-embedded section of rat spleen tissue. Heat media
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000177098-SCN4B/tissue/cerebral+cortex#imid_4551952
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000177098-SCN4B/tissue/colon#imid_4551862
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000177098-SCN4B/tissue/liver#imid_4551863
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000177098-SCN4B/tissue/kidney#imid_4551872
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SCN4B IHC & IF
🧪Run adrenal gland first and assess its glandular cells for SCN4B staining (HPA: adrenal gland glandular cells, Medium). Use adipose tissue as the negative tissue (HPA: adipocytes, Not detected); on the adrenal slide, non-glandular cells should provide a pale or unstained background comparison, though their SCN4B status is unreported.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for SCN4B; 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, a rabbit IgG isotype control matched to the primary antibody’s class, and SCN4B knockout material as a biological negative (selected IHC caption: rabbit primary). Check endogenous peroxidase and biotin background in the adrenal section because the documented detection uses a biotin-based complex and DAB (selected IHC caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A06453-1 paraffin-section caption does not state the fixative. Start with heat retrieval in citrate buffer at pH 6 for 20 minutes (selected IHC caption), while treating retrieval dependency as unestablished; the supplied evidence does not establish that frozen sections or IF are easier (HPA subcellular: no ICC-IF image cell lines). No adrenal-specific artefact is reported; assess glandular background with the stated controls (HPA: adrenal gland glandular cells, Medium).
HPA tissue IHC evidence for SCN4B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SCN4B IHC Tips
Troubleshoot SCN4B staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before interpreting chromogenic signal.
What retrieval should I start with if SCN4B staining is weak?
Use heat-mediated citrate buffer at pH 6 for antigen retrieval (datasheet A06453-1). The selected tissue image used this retrieval for 20 minutes before overnight incubation with the catalog antibody at 1 µg/ml and 4°C (caption A06453-1). If staining remains weak, compare retrieval duration on adjacent sections while keeping antibody concentration and DAB development constant, and inspect morphology for heat damage (standard IHC practice). Include a section processed without primary antibody to distinguish retrieval-associated background from specific signal, and judge any gain against membrane-associated staining and the expected cell distribution (standard IHC practice; UniProt Q8IWT1 topology and tissue specificity).
How should I troubleshoot fixation-related loss of SCN4B signal?
The selected caption describes a paraffin-embedded section but does not state its fixative, so SCN4B-specific sensitivity to fixation is unknown (caption A06453-1). Record the actual fixative and fixation duration for each specimen, then compare sections with matched retrieval and detection conditions when troubleshooting variable staining (standard IHC practice). Use the reported citrate pH 6 retrieval for 20 minutes as the initial processing reference, without treating it as evidence for any fixation condition (caption A06453-1). Examine nuclear and tissue morphology alongside signal intensity, and avoid assigning a fixation effect when section thickness, antibody incubation, or DAB development also differs (standard IHC practice).
Should I expect membrane or cytoplasmic SCN4B staining?
SCN4B is annotated at the cell membrane, with an extracellular region at residues 31–162, a transmembrane segment at 163–183, and a cytoplasmic region at 184–228 (UniProt Q8IWT1 topology). Evaluate membrane-associated staining first, including whether it outlines cells consistently in well-preserved tissue (UniProt Q8IWT1 subcellular location; standard IHC practice). HPA reports variable cytoplasmic staining across tissues and labels its tissue IHC data Approved while noting low consistency with RNA expression (HPA tissue IHC). Treat diffuse cytoplasmic DAB as an observation requiring independent support, especially where cell boundaries are unclear, rather than as proof of the annotated membrane location (HPA tissue IHC; UniProt Q8IWT1 subcellular location).
Could epitope position or SCN4B isoforms explain discordant staining?
SCN4B has 3 annotated isoforms, but the supplied evidence does not identify which sequence the catalog antibody recognizes (UniProt Q8IWT1 isoforms; caption A06453-1). Its extracellular region spans residues 31–162 and contains an Ig-like domain and glycosylation sites at 45, 71, and 113 (UniProt Q8IWT1 topology, domains, and glycosylation). Check the antibody's documented immunogen or mapped epitope before attributing differences between specimens to isoform expression or epitope masking (standard IHC practice). If that information becomes available, compare epitope position with isoform sequences and repeat staining under matched retrieval conditions; staining alone cannot resolve those explanations (standard IHC practice).
How can IF help check a disputed IHC localisation?
Use IF as a separate validation experiment and multiplex SCN4B with a validated marker for the cell population under study; neuronal cells are among the HPA positive populations, and dorsal root ganglia have high reported expression (HPA tissue IHC; UniProt Q8IWT1 tissue specificity). Choose spectrally separated fluorophores, favoring a longer-wavelength channel when the specimen has strong autofluorescence, and include single-label and no-primary controls (standard IF practice). Set permeabilisation according to the mapped epitope: an intracellular epitope generally requires access across the membrane, whereas an extracellular epitope may be tested without permeabilisation (UniProt Q8IWT1 topology; standard IF practice). The supplied evidence does not establish this antibody's IF performance or epitope location, so verify both before interpreting colocalisation (caption A06453-1).
What should I check when SCN4B DAB staining appears diffuse?
Run no-primary and secondary-only controls to identify detection-system signal, and inspect whether DAB accumulates at folds, section edges, or damaged areas (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, and a streptavidin-biotin complex with DAB (caption A06453-1). Confirm that the general chromogenic workflow includes an appropriate endogenous peroxidase block, and assess endogenous biotin when using biotin-based detection (standard IHC practice). Reduce nonspecific staining by optimizing blocking, washing, antibody concentration, and DAB development one variable at a time, while preserving a matched positive section (standard IHC practice).
How should I score SCN4B IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before comparing specimens, because SCN4B is membrane annotated while HPA reports variable cytoplasmic tissue staining (UniProt Q8IWT1 subcellular location; HPA tissue IHC). Report the percentage of positive cells and a predefined intensity scale, or calculate an H-score from those same categories (standard IHC practice). For sparse positive populations, report positive-cell density per mm² of evaluable tissue and exclude folds and necrotic areas by a consistent rule (standard IHC practice). Normalize each result to the counted cells or evaluable tissue area, and compare sections processed with matched retrieval, antibody incubation, and DAB development (standard IHC practice; caption A06453-1).
When is a positive SCN4B stain convincing rather than artefactual?
A convincing result combines reproducible cell-associated staining with an appropriate cell population and compartment: SCN4B is membrane annotated, and reported high expression includes dorsal root ganglia (UniProt Q8IWT1 subcellular location and tissue specificity). HPA reports cytoplasmic staining in many tissues but also low consistency between antibody staining and RNA expression, so cytoplasmic positivity needs additional support (HPA tissue IHC). Suspect artefact when DAB is confined to section edges or necrosis, tracks tissue damage, or persists in a no-primary control; assess endogenous peroxidase during that review (standard IHC practice). Compare adjacent sections and independent evidence before calling unexpected cells positive, particularly when their staining lacks a clear cell boundary (standard IHC practice).
Boster reagents
Best SCN4B / Sodium channel regulatory subunit beta-4 IHC Antibodies
A06453-1 has real SCN4B IHC images from human renal cancer and rat spleen paraffin sections; ICC and IHC-F are listed applications (catalog: IHC image captions, applications).
Real IHC data
Anti-SCN4B Antibody ®
Cat # A06453-1
A06453-1 will render with its human renal cancer paraffin-section IHC figure (catalog: card image caption). An additional caption documents rat spleen paraffin-section IHC; Human, Mouse and Rat reactivity is listed, but no IF image is supplied (catalog: IHC image captions, reactivity, IF image alts).
Which to pick: For tissue IHC, pick A06453-1: its figure documents 1 μg/ml on a human renal cancer paraffin section after citrate pH 6 retrieval; the fixative is unreported (catalog: card image caption). For IF/ICC, A06453-1 lists ICC and IHC-F applications and an IF dilution of 0.5–1 μg/ml, but supplies no IF image (catalog: applications, IF dilution, IF image alts). For cross-species work, A06453-1 lists Human, Mouse and Rat reactivity, with tissue IHC captions for human and rat only; it is rabbit-hosted, and clonality is unreported (catalog: reactivity, IHC image captions, host, clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IWT1 (SCN4B_HUMAN, Sodium channel regulatory subunit beta-4).
- Human Protein Atlas. SCN4B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SCN4B subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SCN4B antibody validation summary (1 antibodies).
- Preserved SCN4B expression is an independent indicator of favorable recurrence-free survival in classical papillary thyroid cancer. PloS one 2018 — PMC5933725.
- Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma. Oncology reports 2026 — PMC12917569.
- SCN4B acts as a metastasis-suppressor gene preventing hyperactivation of cell migration in breast cancer. Nature communications 2016 — PMC5150224.
- Correlating obstructive sleep apnea and lung adenocarcinoma using hub gene signatures and molecular mechanisms. Discover oncology 2026 — PMC13338089.
- PubMed PMID:12930796 — UniProt-cited evidence.
- PubMed PMID:16554811 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SCN4B (Sodium channel regulatory subunit beta-4). Expected localisation Cytoplasm observed (HPA tissue IHC); membrane annotated (UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A06453-1). Validated IHC antibody: A06453-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human renal cancer tissue; fixative not specified (datasheet A06453-1); 2. Fixation: Image fixative and duration unreported (datasheet A06453-1); 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: Citrate pH 6 , 20 min (datasheet A06453-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06453-1); 8. Primary antibody: Rabbit anti-SCN4B, 0.5-1μg/ml (datasheet A06453-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06453-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (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 A06453-1); Caveat: IHC staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; map the epitope to the extracellular or cytoplasmic region (UniProt); Regulation: Higher expression in dorsal root ganglia (UniProt). Evidence: UniProt Q8IWT1, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A06453-1, PMC5150224. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SCN4B/immunohistochemistry-ihc-guide.