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SEPHS1 / Zincore component SEPHS1 · IHC design guide
Design Immunohistochemistry for SEPHS1
Plan chromogenic IHC for SEPHS1 in paraffin sections using general nuclear staining as the tissue benchmark (HPA tissue IHC). The guide highlights high staining in adipocytes from adipose tissue (HPA tissue IHC) and a catalog antibody with human paraffin section examples (datasheet A10102-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SEPHS1 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 tissue (HPA tissue IHC) | |
| Staining pattern | Nuclei across diverse tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10102-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Heart muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10102-1) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Rises until G2/M, then declines (UniProt) | |
| Isoform / epitope | Four isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended SEPHS1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A10102-1). The published IHC protocols below cover liver tissue, cartilage, and mouse paraffin sections (PMC7903722; PMC8828855; PMC8583877).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A10102-1) |
| Fixation | Image fixative and duration unreported (datasheet A10102-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A10102-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10102-1) |
| Primary antibody | Rabbit anti-SEPHS1, 2-5 µg/ml (datasheet A10102-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10102-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10102-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SEPHS1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A10102-1); assess published retrieval conditions when adapting another antibody (PMC8583877).
Section 2
What Is the Expected SEPHS1 Staining Pattern?
SEPHS1 should show predominantly nuclear staining across many cell types in paraffin-section IHC (HPA: general nuclear expression; low tissue specificity). UniProt also lists cytoplasm, cell membrane, nuclear membrane and chromosome locations, so a minor signal outside the nucleus is possible (UniProt P49903: subcellular locations). SEPHS1 has no transmembrane segment (UniProt P49903: topology). HPA rates its tissue IHC evidence Enhanced, with medium agreement between staining and RNA data pending external verification (HPA: reliability).
| Distinct nuclear chromogen in adrenal glandular cells or cerebral cortex neurons (HPA: High in both cell types). | This fits the expected compartment and observed positive cells (HPA: general nuclear expression; High in both). Judge intensity against controls on the same run; High describes HPA's observed staining, not a guaranteed intensity in every specimen. |
| Strong staining confined to cell borders, with little nuclear staining (HPA: general nuclear expression). | The distribution conflicts with the predominant tissue IHC pattern (HPA: general nuclear expression). UniProt lists cell membrane localisation, so do not reject a minor border signal solely on location; assess the nuclear pattern and controls together (UniProt P49903: subcellular locations). |
| Prominent chromogen in cardiomyocytes but absent from adjacent expected positive cells (HPA: cardiomyocytes Not detected). | Check cross-reactivity and endogenous detection activity before calling this SEPHS1. HPA did not detect cardiomyocyte staining; that observation is a comparator, not proof that every heart specimen must be negative (HPA: heart muscle IHC). |
| Diffuse colour across nuclei, cytoplasm and empty tissue spaces, with poor cell boundaries. | Treat the field as background until a control run shows specific localisation. Review blocking, wash stringency and detection controls as general IHC practice; diffuse colour alone does not establish SEPHS1 expression (HPA: general nuclear expression). |
| No nuclear signal in a well-preserved adrenal gland section (HPA: glandular cells High). | First check whether the positive control and detection reagents worked. HPA's High result makes adrenal glandular cells a useful comparator, but its Enhanced rating still has medium RNA agreement pending external verification (HPA: tissue IHC reliability). |
💡Expected SEPHS1 appearanceCall a convincing positive result when identifiable glandular or neuronal cells show clear, predominantly nuclear staining, with intensity judged against run controls (HPA: general nuclear expression; High in adrenal glandular cells and cortical neurons); diffuse field-wide colour or dominant cardiomyocyte staining is suspect (HPA: cardiomyocytes Not detected).
| Cell and tissue context (HPA: tissue IHC). | HPA reports High staining in adipocytes, respiratory epithelial cells, adrenal and duodenal glandular cells, cortical neurons and colonic endothelial cells. It reports Low staining in hippocampal and caudate glia (HPA: tissue IHC); choose a comparator with the relevant cell type. |
| IHC evidence strength (HPA: antibody validation). | HPA037645 and HPA062118 are both rated Enhanced for IHC (HPA: antibody records). HPA describes tissue staining versus RNA agreement as medium and pending external verification (HPA: reliability); interpret an isolated unexpected pattern cautiously. |
| Compartment and topology (UniProt P49903; HPA: tissue IHC). | Nuclear staining has the strongest tissue-pattern support (HPA: general nuclear expression). UniProt also lists cytoplasm and membranes, but reports no transmembrane segment (UniProt P49903: subcellular locations; topology); location alone cannot validate a membrane-dominant stain. |
| Isoforms and cell cycle (UniProt P49903). | Four isoforms are listed, and expression rises through the cell cycle until G2/M before decreasing (UniProt P49903: isoforms; tissue specificity note). Without antibody epitope information, neither an isoform-specific staining pattern nor a fixed intensity threshold can be assigned. |
| IF/ICC Q: Should the IHC pattern transfer directly? | A: Use nucleoplasmic signal as a qualified comparison: HPA calls its ICC-IF location nucleoplasm, uncertain (HPA: subcellular summary). HPA037645 is Uncertain for ICC, and HPA062118 has no ICC rating (HPA: antibody records); this IHC section provides no IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control lacks nuclear staining (HPA: general nuclear expression). | The IHC run may have failed; absence alone cannot identify a SEPHS1-specific cause. | Check section integrity, detection reagents and the assay's antigen-retrieval settings as general IHC practice. Repeat with a known positive cell population, such as adrenal glandular cells (HPA: High), before scoring study tissue. |
| Nuclear stain is present but weak in a purported positive tissue. | The sampled cell type may have lower observed staining: hippocampal and caudate glia are Low (HPA: tissue IHC). | Identify the actual cell population, then compare it with a matched positive control. Do not apply a High-tissue intensity expectation to glia reported Low by HPA (HPA: tissue IHC). |
| Signal is strongest at cell borders with sparse nuclear stain. | The pattern differs from HPA's general nuclear tissue profile, although UniProt lists cell membrane localisation (HPA: tissue IHC; UniProt P49903: subcellular locations). | Recheck morphology and detection controls; assess whether a clear nuclear component exists. Avoid assigning the border-only pattern to SEPHS1 from compartment alone. |
| Chromogen appears in cardiomyocytes (HPA: Not detected). | Cross-reactivity or endogenous detection activity is possible; HPA reports no cardiomyocyte staining (HPA: heart muscle IHC). | Use appropriate reagent-omission and detection controls as general IHC practice. Compare with a known positive cell type before treating the cardiomyocyte signal as target-specific. |
| Diffuse background obscures nuclei and cell boundaries. | Insufficient blocking, washing or detection control can obscure localisation in chromogenic IHC (general IHC practice). | Review those steps and compare control sections processed together. Score SEPHS1 only where cell-level nuclear staining is distinguishable from background (HPA: general nuclear expression). |
| IF/ICC shows no clear nucleoplasmic enrichment. | HPA's nucleoplasm assignment is uncertain, and its antibody records give ICC as Uncertain or unrated (HPA: subcellular summary; antibody records). | Interpret the IF result within its separate guide and controls. Do not use the IHC Enhanced rating as proof of ICC performance (HPA: antibody validation). |
Experiment evidence
IHC & IF images showing SEPHS1 staining
A10102-1 · IHCIHC analysis of Selenophosphate Synthetase 1/SEPHS1 using anti-Selenophosphate Synthetase 1/SEPHS1 antibody (A10102-1). Selenophosphate Synt
A10102-1 · IHCIHC analysis of SEPHS1 using anti-SEPHS1 antibody (A10102-1). SEPHS1 was detected in a paraffin-embedded section of human liver cancer tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000086475-SEPHS1/tissue/cerebral+cortex#imid_8994026
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000086475-SEPHS1/tissue/colon#imid_8989128
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000086475-SEPHS1/tissue/liver#imid_8989113
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000086475-SEPHS1/tissue/kidney#imid_8989130
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037645; green = SEPHS1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037645; green = SEPHS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SEPHS1 IHC & IF
🧪Run cerebral cortex first and look for staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). Run heart muscle as the negative tissue (HPA: Not detected in cardiomyocytes); on the cortex slide, assess cells without specific staining as internal negatives against the no-primary control, without assuming a particular cell type is negative (standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SEPHS1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (uncertain) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control, and SEPHS1 knockout material or a peptide-block control if the immunizing peptide is available (selected-SKU caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase for DAB detection and check brain tissue for background pigment (selected-SKU caption: brain tissue and HRP/DAB detection; standard IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C, but does not establish that retrieval is required (selected-SKU tissue-IHC caption). The fixative and any target-specific fixation window or fixation effect are unreported (selected-SKU tissue-IHC caption). No matched frozen-section or IF comparison establishes an easier format; brain pigment can complicate chromogenic scoring, so assess it alongside the controls (supplied evidence: no matched comparison; standard IHC practice).
HPA tissue IHC evidence for SEPHS1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SEPHS1 IHC Tips
Troubleshoot SEPHS1 staining in paraffin sections by checking retrieval, compartment, cell type, controls, and scoring before interpreting chromogenic signal.
Which retrieval condition should I start with for SEPHS1 paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for SEPHS1 paraffin sections (datasheet A10102-1). The selected image used this condition with 2 μg/ml primary antibody overnight at 4°C, so keep those conditions consistent while evaluating retrieval (caption A10102-1). Compare matched sections processed with the same heating and cooling times, and score nuclear signal separately from background (standard IHC practice; HPA: general nuclear expression). If staining is weak, test another retrieval buffer or heating duration on adjacent sections and record any change in morphology (standard IHC practice). The caption does not report a fixative, so do not treat its retrieval result as proof of performance under every fixation condition (caption A10102-1).
How should I troubleshoot weak staining when fixation details are incomplete?
Target-specific fixation sensitivity for SEPHS1 is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (caption A10102-1). Record the actual fixative and fixation duration for each specimen before comparing staining intensity across batches (standard IHC practice). On matched sections, hold retrieval at EDTA pH 8.0 and the primary incubation at 2 μg/ml overnight at 4°C while assessing fixation-related differences (datasheet A10102-1; caption A10102-1). Check nuclear detail, tissue preservation, and background alongside signal because damaged morphology makes intensity comparisons unreliable (standard IHC practice). Neither the reported tissue staining nor SEPHS1 topology establishes a target-specific response to fixation (HPA: tissue IHC; UniProt P49903 topology).
Where should convincing SEPHS1 chromogenic staining appear?
Prioritize interpretable nuclear staining because tissue IHC reports general nuclear expression, while the subcellular summary places SEPHS1 in the nucleoplasm with uncertainty (HPA: tissue IHC; HPA: subcellular). UniProt also lists chromosome, cytoplasm, cell membrane, and nuclear membrane locations, so document convincing nonnuclear patterns separately rather than merging them into a nuclear score (UniProt P49903 subcellular). Assess localization within intact cells using a counterstain and adjacent morphology; diffuse staining over damaged tissue is difficult to assign to a compartment (standard IHC practice). Compare cellular patterns with a positive reference: cerebral-cortex neuronal cells are reported high, whereas hippocampal glial cells are reported low (HPA: tissue IHC). Recheck unexpected membrane-dominant staining with controls because SEPHS1 has no annotated transmembrane segment (UniProt P49903 topology).
Could isoforms or epitope accessibility explain discordant SEPHS1 staining?
SEPHS1 has 4 listed isoforms, but the supplied antibody caption gives no epitope sequence or isoform coverage (UniProt P49903 isoforms; caption A10102-1). Therefore, do not assign a negative cell population to one isoform without independent epitope information or an orthogonal measurement (standard IHC interpretation). The record lists an N-acetylserine at residue 2 and no annotated glycosylation sites; neither fact establishes whether this antibody recognizes a modified region (UniProt P49903 modified residues; UniProt P49903 glycosylation). Compare adjacent sections under the documented EDTA pH 8.0 retrieval condition before attributing weak staining to epitope masking (datasheet A10102-1; standard IHC practice). Preserve the distinction between an inaccessible epitope and absent protein when reporting negative IHC results (standard IHC interpretation).
How can IF help check a disputed SEPHS1 IHC pattern?
Use IF as a separate localization check: HPA reports nucleoplasmic SEPHS1 with uncertainty and lists ICC/IF images, but the supplied catalog-antibody evidence is for paraffin-section IHC (HPA: subcellular; caption A10102-1). Multiplex with a marker of the expected cell type, such as a neuronal marker when examining cerebral cortex, where neuronal staining is reported high (HPA: cerebral cortex neuronal cells; standard IF practice). Choose fluorophores after checking tissue autofluorescence in unstained controls, and separate channels sufficiently to assess bleed-through (standard IF practice). If testing an intracellular epitope, permeabilize appropriately for nuclear access; the catalog antibody's epitope position is not supplied (UniProt P49903 subcellular; caption A10102-1). Do not transfer the paraffin-image retrieval or incubation conditions directly to IF (caption A10102-1; standard IF practice).
What should I check when SEPHS1 DAB staining is diffuse?
First compare a no-primary control with the stained section to distinguish detection-system background from antibody-dependent signal (standard IHC practice). The documented section used 10% goat-serum blocking, a peroxidase-conjugated secondary antibody, and DAB development (caption A10102-1). Include an endogenous-peroxidase block and assess whether its omission explains diffuse or cell-associated brown signal; this is a general chromogenic IHC step, not SEPHS1-specific evidence (standard IHC practice). If background persists, review primary concentration against the documented 2 μg/ml, wash consistency, and section drying or edge effects (caption A10102-1; standard IHC practice). Interpret faint diffuse cytoplasmic color cautiously when it obscures the reported general nuclear pattern (HPA: tissue IHC).
How should I quantify SEPHS1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine a nuclear scoring rule because HPA describes general nuclear tissue staining, and record cytoplasmic or membrane signal separately (HPA: tissue IHC; UniProt P49903 subcellular). For comparable fields, report the percentage of positive intact cells and an H-score calculated from the fractions at intensity grades 0–3 (standard IHC quantification). Normalize counts to the number of evaluable cells of the same cell type, or report positive-cell density per mm² of viable tissue when area is the denominator (standard IHC quantification). Keep retrieval, detection development, counterstaining, and image thresholds consistent across sections (standard IHC practice). Because HPA reports low tissue RNA specificity, a single positive field does not establish tissue specificity (HPA: tissue IHC RNA specificity).
How do I distinguish true SEPHS1 staining from artefact?
Favor crisp staining assigned to intact nuclei and plausible cell types: HPA reports general nuclear expression and high staining in cerebral-cortex neuronal cells (HPA: tissue IHC). Treat strong staining confined to cut edges, folds, or necrotic areas as suspect and compare with adjacent intact tissue (standard IHC practice). Check no-primary and peroxidase-block controls when brown deposits appear in unexpected compartments, since DAB with peroxidase detection can reveal endogenous enzyme activity (caption A10102-1; standard IHC practice). A membrane-only interpretation needs additional validation because UniProt lists membrane localization but no transmembrane segment, while HPA's principal tissue pattern is nuclear (UniProt P49903 subcellular; UniProt P49903 topology; HPA: tissue IHC). Report discordance explicitly rather than scoring artefactual deposits as positive cells (standard IHC interpretation).
Boster reagents
Best SEPHS1 / Zincore component SEPHS1 IHC Antibodies
The IHC-validated anti-SEPHS1 antibody has paraffin-section images from rat brain, human liver, and human liver cancer; no IF/ICC image is supplied (catalog: A10102-1 image captions).
Real IHC data
Anti-Selenophosphate synthetase 1/SEPHS1 Antibody ®
Cat # A10102-1
A10102-1 will render with an IHC image from a paraffin-embedded rat brain section (catalog: A10102-1 figure caption). Its additional IHC captions show paraffin-embedded human liver and liver cancer sections; listed reactivity is human, mouse, and rat (catalog: A10102-1 image captions and reactivity).
Which to pick: Choose A10102-1 for paraffin-section IHC: its captions document EDTA retrieval at pH 8.0 and 2 µg/ml primary antibody in the pictured rat and human sections; the fixative is unreported (catalog: A10102-1 IHC captions). For cross-species work, it is listed as a rabbit polyclonal reactive with human, mouse, and rat, though the supplied IHC captions show only human and rat tissue (catalog: A10102-1 host, dilution_raw, reactivity, and IHC captions). No IF/ICC option is supported by the supplied application list or images (catalog: A10102-1 applications and if_image_alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49903 (SPS1_HUMAN, Zincore component SEPHS1).
- Human Protein Atlas. SEPHS1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SEPHS1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. SEPHS1 antibody validation summary (2 antibodies).
- SEPHS1 promotes SMAD2/3/4 expression and hepatocellular carcinoma cells invasion. Experimental hematology & oncology 2021 — PMC7903722.
- Selenophosphate synthetase 1 deficiency exacerbates osteoarthritis by dysregulating redox homeostasis. Nature communications 2022 — PMC8828855.
- Identification of Signaling Pathways for Early Embryonic Lethality and Developmental Retardation in Sephs1(-/-) Mice. International journal of molecular sciences 2021 — PMC8583877.
- Identification and Validation of a Nine-Gene Amino Acid Metabolism-Related Risk Signature in HCC. Frontiers in cell and developmental biology 2021 — PMC8452960.
- PubMed PMID:7665581 — UniProt-cited evidence.
- PubMed PMID:20471958 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SEPHS1 (Zincore component SEPHS1). Expected localisation Nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10102-1). Validated IHC antibody: A10102-1 at 2-5 µg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A10102-1); 2. Fixation: Image fixative and duration unreported (datasheet A10102-1); 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: EDTA pH 8.0 (datasheet A10102-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10102-1); 8. Primary antibody: Rabbit anti-SEPHS1, 2-5 µg/ml (datasheet A10102-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10102-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10102-1); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: Four isoforms; check antibody epitope coverage (UniProt); Regulation: Rises until G2/M, then declines (UniProt). Evidence: UniProt P49903, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A10102-1, PMC7903722, PMC8828855. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SEPHS1/immunohistochemistry-ihc-guide.