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ZSCAN16 / Zinc finger and SCAN domain-containing protein 16 · IHC design guide
Design Immunohistochemistry for ZSCAN16
Plan chromogenic IHC on paraffin sections for ZSCAN16 using its general nuclear tissue pattern (HPA tissue IHC). Start with the catalog antibody at 1:100–1:300 (datasheet), and compare high-staining breast glandular cells with liver cholangiocytes where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ZSCAN16 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 | General nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across diverse cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10780-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Liver+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 0 annotated isoforms; one chain, residues 1–348 (UniProt) |
Section 1
Recommended ZSCAN16 IHC & IF Protocols
The catalog antibody uses Tris-EDTA pH 9.0 retrieval (datasheet: A10780-1). One published ZSCAN16 IHC protocol is listed below (PMC11546084: IHC methods).
| Sample | Paraffin-embedded human Breast cancer tissue; fixative not specified (datasheet A10780-1) |
| Fixation | Image fixative and duration unreported (datasheet A10780-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A10780-1); 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-ZSCAN16, 1:100-1:300 (datasheet A10780-1) |
| 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 | ZSCAN16-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 9.0 for the catalog antibody (datasheet: A10780-1); the published protocol used microwave retrieval in sodium citrate (PMC11546084: IHC methods).
Section 2
What Is the Expected ZSCAN16 Staining Pattern?
ZSCAN16 is a nuclear protein with no transmembrane segment (UniProt Q9H4T2: subcellular location and topology). In IHC, expect nuclear staining across multiple cell types, including glandular, hematopoietic, respiratory epithelial and neuronal cells scored High by HPA (HPA: tissue IHC). HPA describes general nuclear expression, with Supported reliability and medium consistency between staining and RNA data (HPA: tissue IHC).
| Distinct nuclear stain in glandular cells of adrenal gland, appendix or breast (HPA: High in these cells). | This matches the reported compartment and provides a useful positive reference (HPA: general nuclear expression; UniProt Q9H4T2: nucleus). Compare nuclei within the same section before treating differences in stain strength as biological differences (standard IHC practice). |
| Predominantly cytoplasmic or membrane-edge stain, with nuclei remaining pale. | That distribution conflicts with the reported nuclear location (UniProt Q9H4T2: nucleus; HPA: general nuclear expression). Review morphology and controls for nonspecific signal or detection artefact (standard IHC practice); the supplied records do not establish an alternative cytoplasmic pool. |
| Strong stain in liver cholangiocytes, smooth muscle cells or spleen red-pulp cells (HPA: Not detected in these cells). | Treat this as discordant with the listed HPA observations, not definitive proof of cross-reactivity: HPA IHC has Supported reliability and medium RNA–staining consistency (HPA: tissue IHC). Check antibody specificity and endogenous detection activity (standard IHC practice). |
| Diffuse stain over tissue, stroma and spaces, with no clear nuclear contrast. | A widespread haze cannot be read as the reported general nuclear pattern (HPA: tissue IHC). Compare a no-primary control and inspect blocking, washing and detection conditions to locate background (standard IHC practice). |
| No nuclear signal in bone-marrow hematopoietic cells or bronchial respiratory epithelial cells (HPA: High in these cells). | This is a failed expected-positive check for that run (HPA: tissue IHC). Verify that the positive tissue, primary antibody and chromogenic detection worked before interpreting a test specimen as ZSCAN16-negative (standard IHC practice). |
💡Expected ZSCAN16 appearanceCall a convincing positive result when chromogenic stain is concentrated in nuclei of a documented High cell population, such as breast glandular cells or cortical neurons (HPA: High; HPA: general nuclear expression); isolated cytoplasmic haze or stain without nuclear contrast is suspect (UniProt Q9H4T2: nucleus; standard IHC practice).
| Compartment and topology (UniProt Q9H4T2: nucleus; no transmembrane segment). | Score nuclear localization against an intact counterstain; a membrane-dominant pattern does not fit this record (UniProt Q9H4T2: nucleus and topology; standard IHC practice). |
| Tissue distribution (HPA: general nuclear expression; low tissue RNA specificity). | High scores occur in several distinct cell populations, while specified cells are Not detected (HPA: tissue IHC). Judge the named cell population, not an entire organ, when selecting positive and discordant references. |
| Strength of IHC evidence (HPA: Supported; medium staining–RNA consistency). | The reported pattern is a guide for interpretation, not an absolute rule for every specimen (HPA: tissue IHC). Antibody HPA007290 is IHC Supported; the supplied antibody record does not label IHC Enhanced (HPA: antibodies). |
| IF/ICC: where should signal appear? (HPA: subcellular ICC-IF). | HPA approves nucleoplasmic localization and lists ICC-IF images from HEK293, U-251MG and U2OS (HPA: subcellular ICC-IF). That localization supports compartment interpretation; this IHC section supplies no IF/ICC protocol. |
| Processing and fixation evidence (UniProt Q9H4T2: chain 1–348; no signal peptide or propeptide). | The record lists no glycosylation sites, modified residues or isoforms (UniProt Q9H4T2). Target-specific fixation sensitivity and antigen-retrieval requirements are unreported in the supplied UniProt and HPA evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-reference tissue has little or no nuclear stain (HPA: bone-marrow hematopoietic cells High). | A weak run, unsuitable retrieval setting or detection failure is possible (standard IHC practice); HPA does not identify a ZSCAN16-specific retrieval condition. | Check counterstain and control performance, then adjust the validated IHC workflow one variable at a time (standard IHC practice). Avoid calling the test tissue negative until the positive reference stains. |
| Stain looks cytoplasmic while expected nuclei are clear (HPA: general nuclear expression). | Off-target or background signal is possible (standard IHC practice); a cytoplasmic ZSCAN16 location is not supported by the supplied record (UniProt Q9H4T2: nucleus). | Inspect focus and cellular boundaries, compare a no-primary control, and reassess the antibody-dependent nuclear signal (standard IHC practice). |
| Chromogen appears in HPA Not detected cell populations (HPA: liver cholangiocytes; smooth muscle cells; spleen red-pulp cells). | Cross-reactivity or endogenous detection activity may contribute (standard IHC practice); HPA's medium RNA–staining consistency limits categorical exclusion (HPA: tissue IHC). | Compare adjacent expected-positive cells and run suitable detection controls before scoring those populations as positive (standard IHC practice). |
| Uniform brown haze obscures nuclear borders. | Nonspecific background from blocking, antibody concentration, washing or detection is possible (standard IHC practice); HPA reports a nuclear pattern (HPA: tissue IHC). | Use a no-primary control to localize background, then refine blocking or washing within the IHC workflow (standard IHC practice). Retain only interpretable nuclear contrast. |
| Some nuclei stain, but the apparent positive cell population is uncertain. | Mixed cell populations and counterstain interpretation can complicate cell-level scoring (standard IHC practice); HPA levels are assigned to named cells (HPA: tissue IHC). | Identify cells by morphology, compare the matching HPA cell entry, and record nuclear staining by cell population rather than assigning one score to the whole tissue (standard IHC practice). |
| A sample differs from the listed HPA intensity despite a working positive control. | The HPA pattern is Supported with medium staining–RNA consistency, so the listed level has interpretive limits (HPA: tissue IHC). | Document compartment, cell identity, intensity and controls; repeat or corroborate a consequential discrepancy before assigning biological meaning (standard IHC practice). |
Experiment evidence
IHC & IF images showing ZSCAN16 staining
A10780-1 · IHCImmunohistochemical analysis of paraffin-embedded human Breast cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196812-ZSCAN16/tissue/cerebral+cortex#imid_2586804
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196812-ZSCAN16/tissue/colon#imid_2586746
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196812-ZSCAN16/tissue/liver#imid_2586747
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000196812-ZSCAN16/tissue/kidney#imid_2586732
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA007290; green = ZSCAN16, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA007290; green = ZSCAN16, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ZSCAN16 IHC & IF
🧪Run breast first: glandular cells should show nuclear staining (HPA: High in breast glandular cells; UniProt Q9H4T2: Nucleus). Use liver cholangiocytes as the negative tissue (HPA: Not detected in liver cholangiocytes); on the breast slide, assess nuclei outside the glandular cells as internal background checks without assuming those cells are ZSCAN16-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Liver (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ZSCAN16 in HEK293, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, immunoglobulin class, concentration and monoclonal or polyclonal format; confirm specificity with a ZSCAN16 knockout sample or peptide competition if the immunizing peptide is available. For chromogenic breast sections, quench endogenous peroxidase and check background; block endogenous biotin if using biotin-based detection.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10780-1 paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). Start retrieval optimization with Tris-EDTA at pH 9.0, the condition reported with 1:200 primary antibody overnight at 4°C (selected-SKU tissue-IHC caption). Frozen sections have no matched protocol here, and IF/ICC is not established as easier; nucleoplasmic IF staining is supported by HPA images (HPA: Nucleoplasm, approved), while breast-specific staining artefacts are unreported.
HPA tissue IHC evidence for ZSCAN16
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 | 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 → |
Section 3
Advanced ZSCAN16 IHC Tips
Troubleshoot ZSCAN16 staining in paraffin sections by checking retrieval, nuclear localisation and controls before comparing signal across specimens.
What retrieval should I try first if nuclear ZSCAN16 staining is weak?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 (datasheet A10780-1). The selected paraffin-section image used that buffer, followed by primary antibody at 1:200 overnight at 4°C (caption A10780-1). If nuclear staining remains weak, adjust heating time within your validated retrieval workflow while keeping matched control sections in the same run (standard IHC practice). Examine tissue morphology alongside signal, since excessive heat can damage sections and make nuclear staining difficult to judge (standard IHC practice). Record the retrieval conditions for each run so that changes in staining can be traced to processing rather than specimen differences (standard IHC practice).
Could fixation explain variable ZSCAN16 staining between paraffin blocks?
ZSCAN16-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A10780-1). Record the fixative and fixation duration for each block, then compare sections processed under similar conditions before assigning biological meaning to intensity differences (standard IHC practice). If archived blocks vary, stain them together with the same retrieval and detection steps and include a control section in each run (standard IHC practice). Uneven preservation or tissue damage should prompt review of morphology before scoring nuclear signal (standard IHC practice). Neither the reported nuclear location nor the absence of annotated glycosylation sites establishes a fixation response (UniProt Q9H4T2).
Where should convincing ZSCAN16 staining appear in a paraffin section?
Expect predominantly nuclear staining: ZSCAN16 is annotated as nuclear, and the subcellular profile places it in the nucleoplasm (UniProt Q9H4T2; HPA subcellular). Assess signal within intact nuclei using the counterstain to define nuclear boundaries, and record cytoplasmic colour separately (standard IHC practice). The tissue profile describes general nuclear expression, with high staining reported in breast glandular cells and bone marrow hematopoietic cells (HPA tissue IHC). A predominantly membranous pattern deserves troubleshooting because the protein has no annotated transmembrane segment (UniProt Q9H4T2). Compare suspicious areas with a negative reagent control and adjacent intact tissue before accepting compartment-specific staining (standard IHC practice).
How can epitope uncertainty affect interpretation of a negative ZSCAN16 result?
The record describes one chain spanning residues 1–348, a SCAN box at 41–123, and no annotated isoforms (UniProt Q9H4T2). Those annotations do not identify the catalog antibody’s epitope or establish whether processing exposes it in every paraffin block (UniProt Q9H4T2; caption A10780-1). When staining is absent, first check retrieval, antibody incubation and detection against a concurrently stained control with expected nuclear signal (standard IHC practice; HPA tissue IHC). Keep epitope uncertainty in the report if those controls work but the specimen remains negative (standard IHC practice). Do not attribute a negative result to a particular ZSCAN16 isoform without additional evidence (UniProt Q9H4T2).
How should I plan IF confirmation of a nuclear IHC pattern?
Use IF as a separate confirmation experiment, with a nuclear counterstain to test the expected nucleoplasmic pattern (HPA subcellular; standard IF practice). Multiplex with a validated marker for the cell population being assessed, and check single-stain controls for channel bleed-through (standard IF practice). Choose a fluorophore channel after examining the specimen’s autofluorescence, especially if broad background obscures nuclear boundaries (standard IF practice). Because ZSCAN16 is nuclear and has no annotated transmembrane segment, permeabilisation should allow antibody access to the nuclear epitope while preserving morphology (UniProt Q9H4T2; standard IF practice). HPA lists ICC/IF images from HEK293, U-251MG and U2OS cells; those images do not establish an IF protocol for the catalog antibody (HPA subcellular; caption A10780-1).
How do I reduce diffuse brown signal without losing nuclear ZSCAN16 staining?
Begin by comparing the stained section with a negative reagent control and checking whether brown signal follows nuclei defined by the counterstain (standard IHC practice; HPA subcellular). For peroxidase-based chromogenic detection, block endogenous peroxidase and assess nonspecific detection-reagent binding as general workflow controls (standard IHC practice). The selected image used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature (caption A10780-1). If background persists, titrate incubation conditions against a control section while preserving nuclear contrast (standard IHC practice). Treat diffuse cytoplasmic or extracellular colour cautiously because the reported ZSCAN16 location is nuclear (UniProt Q9H4T2; HPA subcellular).
How should I score ZSCAN16 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define positive staining as signal within intact nuclei, consistent with the reported nucleoplasmic location (HPA subcellular; standard IHC practice). For a specified cell population, report the percentage of positive nuclei and an intensity-based H-score on its conventional 0–300 scale (standard IHC practice). Normalise counts to the number of evaluable nuclei in that population, and record the sampled area if reporting positive nuclei per mm² (standard IHC practice). Keep retrieval, detection, imaging and scoring thresholds consistent across compared sections (standard IHC practice). Note the cell population and control performance beside each score, since the tissue profile reports general nuclear expression with variable cell-level detection (HPA tissue IHC).
When is apparent ZSCAN16 positivity more likely to be artefact?
Give greatest weight to staining in intact nuclei, matching the nuclear and nucleoplasmic annotations (UniProt Q9H4T2; HPA subcellular). Check whether the labelled cell population is plausible: HPA reports high staining in breast glandular cells but no detection in liver cholangiocytes or smooth muscle cells (HPA tissue IHC). Edge-only colour, necrotic areas and staining in a negative reagent control warrant review for processing or detection artefact (standard IHC practice). In a peroxidase-DAB workflow, investigate endogenous enzyme activity when brown colour persists in the relevant negative control (standard IHC practice). HPA rates its tissue staining evidence as Supported with medium consistency against RNA expression, so resolve discordant specimens with matched controls before drawing conclusions (HPA tissue IHC).
Boster reagents
Best ZSCAN16 / Zinc finger and SCAN domain-containing protein 16 IHC Antibodies
A10780-1 lists human and mouse reactivity and IF (catalog: applications and reactivity); its real IHC image shows paraffin-embedded human breast cancer (IHC image caption).
Real IHC data
Anti-ZSCAN16 Antibody
Cat # A10780-1
A10780-1 has an IHC image from paraffin-embedded human breast cancer (IHC image caption). A10780-1 also lists IF and human/mouse reactivity, but supplies no IF image or mouse sample image (catalog: applications, reactivity and image captions).
Which to pick: For paraffin-section IHC, choose A10780-1: its image documents Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 overnight; the fixative is unreported (IHC image caption). For IF, A10780-1 lists a 1:200–1:1000 range, but has no IF image and does not list ICC (catalog: applications, IF dilution and IF image captions). For cross-species work, A10780-1 lists human and mouse reactivity, although its IHC image shows only human tissue; the host is rabbit and clonality is unreported (catalog: reactivity, host and clone; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H4T2 (ZSC16_HUMAN, Zinc finger and SCAN domain-containing protein 16).
- Human Protein Atlas. ZSCAN16 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ZSCAN16 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. ZSCAN16 antibody validation summary (1 antibodies).
- ZSCAN16 expedites hepatocellular carcinoma progression via activating TBC1D31. Cell division 2024 — PMC11546084.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ZSCAN16 (Zinc finger and SCAN domain-containing protein 16). Expected localisation General nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10780-1). Validated IHC antibody: A10780-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Liver. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Breast cancer tissue; fixative not specified (datasheet A10780-1); 2. Fixation: Image fixative and duration unreported (datasheet A10780-1); 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: Tris-EDTA pH 9.0 (datasheet A10780-1); 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-ZSCAN16, 1:100-1:300 (datasheet A10780-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one chain, residues 1–348 (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q9H4T2, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A10780-1, PMC11546084. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ZSCAN16/immunohistochemistry-ihc-guide.