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ZNF703 / Zinc finger protein 703 · IHC design guide
Design Immunohistochemistry for ZNF703
Plan ZNF703 paraffin-section IHC around widespread nuclear tissue staining (HPA tissue IHC). This guide covers fixation consistency, controls using skin fibrohistiocytic cells with high staining, and interpretation across cell types (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ZNF703 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 tissue staining (HPA tissue IHC); cytoplasm annotated (UniProt) | |
| Staining pattern | Widespread nuclear staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Skin+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Skin fibrohistiocytic cells stain high; other cell types vary (HPA tissue IHC) | |
| Regulation | Cell-type-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | No isoforms annotated; chain spans residues 2–590 (UniProt) |
Section 1
Recommended ZNF703 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published ZNF703 IHC protocols for breast and ovarian specimens (PMC8889678; PMC3753350; PMC7693506).
| Sample | Paraffin-embedded rat pancreas tissue; fixative not specified (datasheet A06570) |
| Fixation | Image fixative and duration unreported (datasheet A06570); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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-ZNF703, 1:50-1:200 (datasheet A06570) |
| 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 | ZNF703-positive staining in fibrohistiocytic cells of skin (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min for nuclear ZNF703 staining (page retrieval rule; UniProt Q9H7S9: nucleus).
Section 2
What Is the Expected ZNF703 Staining Pattern?
ZNF703 should appear predominantly nuclear in paraffin-section IHC, with possible cytoplasmic signal (HPA: ubiquitous nuclear expression; UniProt Q9H7S9: nucleus and cytoplasm). Skin fibrohistiocytic cells show high staining; several other listed cell populations show medium staining (HPA tissue IHC). Interpret these expectations with the reported Supported reliability, which reflects medium consistency between antibody staining and RNA data (HPA tissue IHC). ZNF703 has no transmembrane segment (UniProt Q9H7S9).
| Discrete nuclear stain in skin fibrohistiocytic cells, with visibly weaker staining in some other cell populations. | This fits the high skin signal and broadly nuclear profile; HPA reports medium signal in selected adipocytes, glandular cells, respiratory epithelium and glia (HPA tissue IHC). Compare cell types within the same section before assigning intensity; HPA’s categories describe its observations, not a universal scoring threshold (HPA tissue IHC). |
| Signal is confined to membranes, luminal material or extracellular deposits, with no convincing nuclear staining. | That compartment pattern conflicts with the observed nuclear profile and the lack of a transmembrane segment (HPA tissue IHC; UniProt Q9H7S9 topology). Check a known-positive cell population and detection controls before calling it ZNF703. Cytoplasmic staining alone deserves review rather than automatic rejection because UniProt also lists cytoplasm (UniProt Q9H7S9). |
| A cell population stains strongly where HPA reports low staining, while expected positive cells stain weakly. | Possible explanations include cross-reactivity, endogenous chromogen-generating activity or a technical mismatch between sections (general IHC practice). Treat the contrast as a reason to investigate, not proof of an artefact: HPA’s low categories are observations, not validated negatives, and its staining-to-RNA consistency is medium (HPA tissue IHC). |
| Brown precipitate is diffuse across tissue compartments or appears similarly in many unrelated cells. | Poorly localized signal can obscure whether nuclei contain genuine stain (general IHC practice). Inspect reagent-only and no-primary controls, morphology and local tissue background before scoring. HPA’s expected pattern is ubiquitous nuclear expression, which means widespread positive cells alone do not establish nonspecific staining (HPA tissue IHC). |
| Skin fibrohistiocytic cells lack nuclear signal despite adequate tissue morphology. | The absent high reference pattern warrants a run-level check (HPA tissue IHC). Review primary-antibody use, retrieval, detection and counterstain with the same run’s controls (general IHC practice). A failed reference does not establish that the specimen lacks ZNF703; HPA reports a supported pattern, not guaranteed positivity in every section (HPA tissue IHC). |
💡Expected ZNF703 appearanceCall a result consistent when intact cells show predominantly nuclear stain, especially high staining in skin fibrohistiocytic cells; isolated membrane or extracellular stain without nuclear signal is suspect (HPA tissue IHC; UniProt Q9H7S9 topology).
| Compartment used for scoring | HPA describes a ubiquitous nuclear IHC pattern, while UniProt lists both nucleus and cytoplasm (HPA tissue IHC; UniProt Q9H7S9). Prioritize interpretable nuclear staining and assess any cytoplasmic signal in its cellular context. |
| Reference cell population | Skin fibrohistiocytic cells are high; adipocytes, adrenal and appendix glandular cells, respiratory epithelial cells and specified neural cells are medium (HPA tissue IHC). HPA also lists low-staining populations, so these are comparison points rather than proven negative controls. |
| Strength of the tissue evidence | The tissue profile is Supported, with medium agreement between antibody staining and RNA expression (HPA tissue IHC). HPA reports tissue-enhanced RNA in skeletal muscle; that RNA category does not specify a skeletal-muscle IHC staining level (HPA tissue IHC). |
| Antibody and application evidence | HPA lists Supported IHC results for HPA023930 and CAB068249, but no ICC validation status for either (HPA antibodies). HPA provides no ICC-IF images or main subcellular location here, so an IF result needs its own controls and interpretation (HPA subcellular). |
| Protein processing and retrieval limits | UniProt lists no signal peptide, propeptide, transmembrane segment or annotated isoform for this record (UniProt Q9H7S9). The supplied sources give no antibody epitope or target-specific fixation effect; choose and verify retrieval conditions from the IHC-validated antibody’s instructions (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in the specimen or the skin reference. | The run may have failed at retrieval, primary-antibody incubation or chromogenic detection (general IHC practice). | Check the run’s positive and no-primary controls, reagent preparation and the IHC-validated antibody’s stated conditions before interpreting absence. Skin fibrohistiocytic cells provide an expected high reference, not an absolute guarantee (HPA tissue IHC). |
| Nuclei appear negative but cytoplasm stains. | UniProt includes cytoplasm, whereas HPA’s tissue pattern is nuclear; either a real compartment difference or nonspecific signal needs assessment (UniProt Q9H7S9; HPA tissue IHC). | Compare nuclear and cytoplasmic signal in a positive reference, inspect no-primary controls and report the observed compartments explicitly (general IHC practice). Do not label cytoplasm alone definitive without further support. |
| Only outlines of cells or extracellular areas are brown. | A membrane- or deposit-only pattern conflicts with the nuclear tissue profile and nonmembrane topology (HPA tissue IHC; UniProt Q9H7S9 topology). | Inspect morphology and no-primary controls, then confirm that a known-positive cell population develops nuclear stain in the same run (general IHC practice). |
| Background masks nuclear detail across the section. | Detection background or inadequate blocking can make a nuclear call unreliable (general IHC practice). | Compare no-primary and detection-only controls; optimize blocking, washing and chromogen development for the assay, then rescore nuclei only where cellular boundaries remain clear (general IHC practice). |
| Unexpected strong staining occurs in an HPA low-staining cell population. | HPA’s low category is not a validated negative; assay background or cross-reactivity is also possible (HPA tissue IHC; general IHC practice). | Confirm the cell identity and compartment, compare controls and a reported high population, and describe the discrepancy without converting the HPA low call into an absolute absence claim (HPA tissue IHC; general IHC practice). |
| An IF/ICC image seems to disagree with the IHC result. | The supplied HPA subcellular record has no ICC-IF images or main-location call, and the listed antibodies have no ICC status here (HPA subcellular; HPA antibodies). | Evaluate IF with its own controls and dedicated guide; use the paraffin-section nuclear pattern as context, without treating IHC support as IF validation (HPA tissue IHC; HPA antibodies). |
Experiment evidence
IHC & IF images showing ZNF703 staining
A06570 · IHCImmunohistochemistry of paraffin-embedded rat pancreas using ZNF703 antibody at dilution of 1:100 .Perform microwave antigen retrieval with
A06570 · IHCImmunohistochemistry of paraffin-embedded human breast cancer using ZNF703 antibody at dilution of 1:100 .Perform microwave antigen retrieva
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000183779-ZNF703/tissue/cerebral+cortex#imid_18630148
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000183779-ZNF703/tissue/colon#imid_18630058
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000183779-ZNF703/tissue/liver#imid_18630067
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000183779-ZNF703/tissue/kidney#imid_18630068
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ZNF703 IHC & IF
🧪Run skin first: fibrohistiocytic cells should stain (HPA: High in skin fibrohistiocytic cells). HPA detects ZNF703 in all 45 scored tissues, so use no-primary and isotype controls instead of a negative tissue; treat neighboring cells as internal negatives only if their signal matches those controls (HPA: no negative tissue listed; standard IHC practice).
Positive control tissue: Skin (Fibrohistiocytic cells, HPA High)
Negative control tissue: None in HPA: ZNF703 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ZNF703; derive a cell-line control from the positive tissue's cell type (Fibrohistiocytic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a nonimmune antibody matched to the primary antibody’s host species, isotype, and clonality; and, if available, ZNF703 knockout material or a validated peptide-block control (standard IHC practice). Quench endogenous peroxidase and distinguish skin pigment from chromogenic signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A06570 rat-pancreas paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption uses microwave retrieval in 10 mM PBS, pH 7.2, but does not establish that retrieval condition as required for skin (selected-SKU tissue-IHC caption). The evidence does not establish that frozen sections or IF/ICC are easier; HPA lists no cell lines with ICC-IF images for ZNF703 (HPA subcellular).
HPA tissue IHC evidence for ZNF703
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 |
|---|---|---|---|---|
| Skin | Fibrohistiocytic cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ZNF703 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ZNF703 IHC Tips
Troubleshoot ZNF703 staining in paraffin sections by checking retrieval, nuclear localisation, controls, and scoring before interpreting chromogenic signal.
How should I retrieve ZNF703 in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA pH 9.0 heat retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Let sections cool in retrieval buffer, then compare staining with a matched section processed in the same run so changes in nuclear signal are interpretable (standard IHC practice). If staining remains weak, test microwave retrieval in 10 mM PBS at pH 7.2 as a fallback; that condition accompanies the 1:100 paraffin rat pancreas image for SKU A06570 (selected-SKU caption). Keep detection and counterstaining constant across the comparison (standard IHC practice). Evaluate nuclear signal against the reported ubiquitous nuclear tissue pattern (HPA: tissue IHC profile).
Could fixation explain weak or uneven ZNF703 staining?
ZNF703-specific fixation sensitivity is unknown because the supplied paraffin rat pancreas caption does not state a fixative (selected-SKU caption). Record the fixative, fixation duration, processing history, and section age for each specimen before comparing staining (standard IHC practice). When investigating weak signal, stain a consistently processed control alongside the affected sections and hold retrieval and antibody concentration constant (standard IHC practice). Excessive or uneven fixation can alter epitope access in paraffin IHC generally, but that effect has not been demonstrated specifically for ZNF703 here (standard IHC practice; selected-SKU caption). Do not infer fixation tolerance from the observed nuclear tissue pattern (HPA: tissue IHC profile).
Should ZNF703 staining be nuclear, cytoplasmic, or both?
Use nuclear staining as the principal tissue IHC readout because the reported profile is ubiquitous nuclear expression (HPA: tissue IHC profile). UniProt also lists cytoplasmic localisation, so document reproducible cytoplasmic signal separately rather than merging it into a nuclear score (UniProt Q9H7S9: subcellular location; standard IHC practice). ZNF703 has no annotated transmembrane segment, making a sharp cell-surface rim an unexpected pattern that warrants scrutiny (UniProt Q9H7S9: topology). Compare the compartment pattern in morphologically intact cells across replicate sections and examine a no-primary control for detection artefacts (standard IHC practice). A nuclear counterstain can define cell boundaries for compartment-specific review (standard IHC practice).
How can epitope choice complicate ZNF703 interpretation?
Check the antibody immunogen or mapped epitope before attributing differences between specimens to ZNF703 biology (standard IHC practice). The supplied record annotates no isoforms and lists a 590-aa protein; absence of an isoform annotation does not establish that every possible variant shares the antibody epitope (UniProt Q9H7S9: length and isoforms). Reported modified residues include acetylserine at position 2, phosphoserine at 252, and methylarginine at 580 (UniProt Q9H7S9: modified residues). If an epitope overlaps one of these sites, test recognition rather than assuming that modification changes staining (standard IHC practice; UniProt Q9H7S9: modified residues). Compare localisation and control staining under the same retrieval conditions (standard IHC practice).
How should I adapt ZNF703 assessment to multiplex IF?
Treat IF as a separate application requiring its own antibody validation; the supplied SKU A06570 image documents paraffin tissue IHC, with fixation unreported (selected-SKU caption). Multiplex ZNF703 with a validated marker for the cell population being assessed, and use a nuclear counterstain to separate nuclear from cytoplasmic signal (standard IF practice; UniProt Q9H7S9: subcellular location). Select fluorophores after checking unstained tissue for autofluorescence, favoring a spectrally separated channel with adequate signal for ZNF703 (standard IF practice). Because ZNF703 has no annotated transmembrane segment and is reported in nucleus and cytoplasm, optimise permeabilisation for intracellular epitope access without assuming a membrane-facing epitope (UniProt Q9H7S9: topology and subcellular location; standard IF practice). Include single-colour and no-primary controls (standard IF practice).
What should I change when chromogenic ZNF703 staining has high background?
Inspect no-primary controls first to distinguish antibody-dependent signal from detection chemistry or tissue pigment (standard IHC practice). In chromogenic IHC, a peroxidase block addresses endogenous enzyme activity, while appropriate protein blocking and washes can reduce nonspecific staining (standard IHC practice). Titrate the catalog antibody around the 1:100 dilution documented for the paraffin rat pancreas image of SKU A06570, keeping retrieval and detection fixed during that comparison (selected-SKU caption; standard IHC practice). Assess whether colour accumulates at section edges, in damaged tissue, or outside intact cells before calling it positive (standard IHC practice). Judge retained signal against the expected nuclear tissue pattern (HPA: tissue IHC profile).
How should I score heterogeneous ZNF703 staining across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population and exclude damaged regions before scoring, using morphology and the same inclusion rules across specimens (standard IHC practice). For nuclear ZNF703, record the percentage of positive eligible nuclei and staining intensity, then calculate an H-score if intensity categories are reproducible (HPA: tissue IHC profile; standard IHC practice). Normalise counts to the number of eligible cells, or report positive-cell density per mm² of evaluated tissue when cell counts are impractical (standard IHC practice). Score cytoplasmic staining separately because UniProt also lists that compartment (UniProt Q9H7S9: subcellular location). Keep retrieval, chromogen development, imaging settings, and thresholds consistent across the comparison (standard IHC practice).
How do I separate genuine ZNF703 positivity from staining artefacts?
Give greatest weight to staining in intact nuclei, consistent with the reported ubiquitous nuclear tissue profile (HPA: tissue IHC profile). Consider cell identity: high staining is reported in skin fibrohistiocytic cells, while low staining is reported in oral mucosa squamous epithelial cells, so interpret intensity within the sampled cell population (HPA: tissue IHC levels). Review isolated membrane rims cautiously because ZNF703 has no annotated transmembrane segment (UniProt Q9H7S9: topology). Edge staining, necrotic regions, and signal retained in a no-primary control can indicate artefact; endogenous peroxidase is a possible source in chromogenic detection (standard IHC practice). Confirm uncertain patterns in independently processed sections before making a biological claim (standard IHC practice).
Boster reagents
Best ZNF703 / Zinc finger protein 703 IHC Antibodies
A06570 is listed for ZNF703 IHC and IF/ICC in human, mouse, and rat (catalog: applications and reactivity); its IHC images show paraffin sections (catalog: IHC image captions).
Real IHC data
Anti-Zinc finger protein 703 ZNF703 Antibody
Cat # A06570
A06570 has IHC images from paraffin-embedded rat pancreas and human breast cancer, each stained at 1:100 (catalog: IHC image captions). It is listed for IF/ICC in human, mouse, and rat, but no IF image is supplied (catalog: applications, reactivity, IF image alts).
Which to pick: Choose A06570 for paraffin-section IHC because its own captions document rat pancreas and human breast cancer staining after microwave retrieval in 10 mM PBS, pH 7.2; the fixative is unreported (catalog: IHC image captions). For IF/ICC, A06570 is listed at 1:50–1:200, without an IF image in the payload (catalog: applications, IF dilution, IF image alts). It is the listed cross-species option for human, mouse, and rat; its host is rabbit and its clonality is unreported (catalog: reactivity, host, clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H7S9 (ZN703_HUMAN, Zinc finger protein 703).
- Human Protein Atlas. ZNF703 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ZNF703 subcellular location (ICC-IF): Highest expression in HEK293: 147.2 nTPM.
- Human Protein Atlas. ZNF703 antibody validation summary (2 antibodies).
- Zinc finger protein 703 induces EMT and sorafenib resistance in hepatocellular carcinoma by transactivating CLDN4 expression. Cell death & disease 2020 — PMC7142083.
- ZNF703 promotes triple-negative breast cancer cells through cell-cycle signaling and associated with poor prognosis. BMC cancer 2022 — PMC8889678.
- Luminal breast cancer cell lines overexpressing ZNF703 are resistant to tamoxifen through activation of Akt/mTOR signaling. PloS one 2013 — PMC3753350.
- ZNF703 promotes tumor progression in ovarian cancer by interacting with HE4 and epigenetically regulating PEA15. Journal of experimental & clinical cancer research : CR 2020 — PMC7693506.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ZNF703 (Zinc finger protein 703). Expected localisation Nuclear tissue staining (HPA tissue IHC); cytoplasm annotated (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A06570 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Skin (Fibrohistiocytic cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat pancreas tissue; fixative not specified (datasheet A06570); 2. Fixation: Image fixative and duration unreported (datasheet A06570); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 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-ZNF703, 1:50-1:200 (datasheet A06570); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Skin fibrohistiocytic cells stain high; other cell types vary (HPA tissue IHC); Isoform / epitope: No isoforms annotated; chain spans residues 2–590 (UniProt); Regulation: Cell-type-dependent expression (HPA tissue IHC). Evidence: UniProt Q9H7S9, HPA tissue IHC · Supported, HPA cell-line RNA · Highest expression in HEK293: 147.2 nTPM, Datasheet A06570, PMC8889678, PMC3753350. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ZNF703/immunohistochemistry-ihc-guide.