OVOL1 / Putative transcription factor Ovo-like 1 · IHC design guide

Design Immunohistochemistry for OVOL1

Plan OVOL1 chromogenic IHC in paraffin sections using the catalog antibody's 1:100–1:300 dilution range (datasheet A11495-1). Assess the reported cytoplasmic tissue staining (HPA tissue IHC) alongside the annotated nuclear location (UniProt), and interpret results cautiously because tissue staining reliability is uncertain (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for OVOL1 (IHC for OVOL1): expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location (UniProt), antibody A11495-1, validated IHC image, and IHC protocol steps — Printable **OVOL1** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location (UniProt), antibody A11495-1, controls and protocol steps. Open the full OVOL1 IHC guide →

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OVOL1 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic tissue staining (HPA tissue IHC); nuclear location (UniProt)
	Staining pattern	Cytoplasmic staining in squamous epithelial cells (HPA tissue IHC)
	Antigen retrieval	Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A11495-1)
	Positive control ⓘ	Cervix+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep paraffin-section fixation consistent across samples (standard IHC practice; not target-specific)
	Caveat	Presumed off-target binding; IHC/RNA concordance is very low (HPA tissue IHC)
	Regulation	Expression varies by tissue (UniProt)
	Isoform / epitope	No isoforms or processing annotated; epitope effects unknown (UniProt)

Section 1

Recommended OVOL1 IHC & IF Protocols

The catalog antibody uses heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet: A11495-1). The published IHC protocols below cover salivary tumors, skin lesions, and pancreatic tissues (PMC12603596; PMC7141138; PMC12304111).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A11495-1)
Fixation	Image fixative and duration unreported (datasheet A11495-1); 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 A11495-1); 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-OVOL1, 1:100-1:300 (datasheet A11495-1)
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	OVOL1-positive staining in squamous epithelial cells of cervix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, including squamous epithelial. No signal in the no-primary control.

💡Decision noteStart with heat-mediated Tris-EDTA at pH 9.0 for the catalog antibody (datasheet: A11495-1). The skin study used pH 6 with a different antibody (PMC7141138).

Section 2

What Is the Expected OVOL1 Staining Pattern?

OVOL1 is annotated as nuclear and has no transmembrane segment (UniProt O14753: subcellular location and topology). In paraffin section IHC, assess signal in squamous epithelial cells of cervix, esophagus, or vagina, and glandular cells of prostate, where HPA reports high staining (HPA: tissue IHC). HPA describes the tissue staining as cytoplasmic but rates its reliability Uncertain because antibody staining and RNA expression have very low consistency and presumed off-target binding was observed (HPA: tissue IHC).

What am I looking at on my slide?
Defined nuclear staining in squamous epithelial cells, with limited surrounding background.	This fits OVOL1's nuclear annotation and the cell types HPA scores High in cervix, esophagus, and vagina (UniProt O14753: subcellular location; HPA: tissue IHC). Treat it as a plausible result requiring validation: the HPA tissue IHC reliability is Uncertain (HPA: tissue IHC).
Cytoplasmic staining dominates, with little or no convincing nuclear signal.	The compartment conflicts with UniProt's nuclear annotation and HPA's approved nucleoplasmic ICC-IF location (UniProt O14753: subcellular location; HPA: subcellular). HPA also reports cytoplasmic tissue staining, but flags that tissue result as Uncertain; cytoplasmic signal alone cannot establish OVOL1 specificity (HPA: tissue IHC).
Strong signal appears mainly outside the expected cells, or in a negative comparison cell population.	Check for cross-reactivity or endogenous detection activity before assigning that signal to OVOL1. HPA scores cervical, esophageal, and vaginal squamous epithelial cells High, while adipocytes in adipose tissue are Not detected (HPA: tissue IHC). Those are cell-specific observations, not a guarantee about every cell in either tissue.
Colour spreads across cells and stroma without clear cell boundaries or a nuclear pattern.	Diffuse staining cannot support a compartment call. Assess background with a no-primary control, then review blocking, washing, and detection conditions as general IHC practice. The need for a clear nuclear call follows the UniProt nuclear annotation; HPA tissue IHC remains Uncertain (UniProt O14753: subcellular location; HPA: tissue IHC).
No signal appears in squamous epithelial cells of an HPA High tissue.	First check section quality, detection, and the antibody's IHC-P conditions as general IHC practice. HPA scores those cells High in cervix, esophagus, and vagina, but its Uncertain reliability means a blank section does not by itself establish absent OVOL1 or a failed assay (HPA: tissue IHC).

💡Expected OVOL1 appearanceA plausible positive is defined nuclear staining in squamous epithelial cells of cervix, esophagus, or vagina, where HPA reports High staining; cytoplasmic-only or diffuse signal is insufficient for a confident OVOL1 call given the conflict between nuclear localisation and Uncertain tissue IHC (UniProt O14753: subcellular location; HPA: tissue IHC and subcellular).

How each factor affects the staining
Compartment agreement	UniProt places OVOL1 in the nucleus, and HPA approves nucleoplasm as the main ICC-IF location (UniProt O14753: subcellular location; HPA: subcellular). HPA tissue IHC instead describes cytoplasmic expression and rates that staining Uncertain, so report the observed compartment explicitly rather than treating either image type as proof of tissue IHC specificity (HPA: tissue IHC).
Cell and tissue choice	HPA scores squamous epithelial cells High in cervix, esophagus, and vagina; prostate glandular cells are also High, while oral mucosal squamous epithelial cells are Medium (HPA: tissue IHC). Assess the named cell population within a section. HPA's Uncertain tissue reliability limits how strongly any one positive or negative comparison can validate the assay (HPA: tissue IHC).
Antibody validation limit	The listed antibody, HPA003984, is Uncertain for IHC and Approved for ICC (HPA: antibodies). HPA attributes its Uncertain tissue assessment to very low agreement between staining and RNA expression and to presumed off-target binding (HPA: tissue IHC). Use morphology, compartment, controls, and independent evidence when interpreting IHC signal.
Topology and processing	OVOL1 has no transmembrane segment, signal peptide, propeptide, or annotated glycosylation sites; its annotated chain spans residues 1–267 (UniProt O14753: topology, processing, glycosylation). These annotations provide no basis to predict a membrane pattern or a shed-product pattern. They also do not establish an antigen retrieval condition or OVOL1-specific fixation sensitivity.
IF/ICC Q&A	Q: What compartment should an IF/ICC result show? A: HPA lists nucleoplasm as the approved main ICC-IF location, consistent with UniProt's nuclear annotation (HPA: subcellular; UniProt O14753: subcellular location). This answers localisation only; IF/ICC procedure selection belongs to its separate guide.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Cytoplasmic signal is stronger than nuclear signal in a candidate positive section.	The observed compartment conflicts with nuclear and approved nucleoplasmic annotations, although HPA describes cytoplasmic tissue staining with Uncertain reliability (UniProt O14753: subcellular location; HPA: subcellular and tissue IHC).	Record nuclear and cytoplasmic scores separately. Check a no-primary control and seek independent evidence before calling cytoplasmic staining specific; these are general IHC interpretation steps.
The expected squamous epithelial cells show no visible chromogenic signal.	HPA scores those cells High in cervix, esophagus, and vagina, but its tissue IHC reliability is Uncertain (HPA: tissue IHC). A technical failure or a weakly reproducible reference pattern remains possible.	Confirm that the section and detection controls worked, then review the chosen antibody's IHC-P instructions, including its stated retrieval and dilution conditions if provided. Do not infer a target-specific fixation effect.
Staining is widespread across cells or connective tissue.	Diffuse background can obscure cell and compartment boundaries; it does not match the discrete nuclear location annotated for OVOL1 (UniProt O14753: subcellular location).	As general IHC practice, compare a no-primary control and review blocking, washing, detection reagent activity, and counterstain before scoring positive cells.
A comparison section stains in adipocytes expected to be negative.	HPA reports adipocytes in adipose tissue as Not detected, but the overall tissue IHC assessment is Uncertain (HPA: tissue IHC). Unexpected staining could reflect nonspecific antibody binding or endogenous detection activity.	Check the cell identity and a no-primary control; localise any residual signal before treating the comparison as evidence about OVOL1. Apply the HPA result to the specified cell population only.
Nuclear and cytoplasmic calls differ between IHC and ICC-IF.	HPA reports cytoplasmic tissue IHC with Uncertain reliability, while its approved ICC-IF location is nucleoplasm (HPA: tissue IHC and subcellular). The records do not identify a mechanism for the difference.	Report each application's observed compartment and validation status separately. For IHC, assess cell type, background controls, and independent evidence before assigning specificity.
A weak signal appears in a tissue expected to be negative from UniProt.	UniProt reports no expression in intestine, peripheral blood lymphocytes, and ovary, while the supplied HPA IHC negatives refer to specific cells in other tissues; those records are not interchangeable (UniProt O14753: tissue specificity; HPA: tissue IHC).	Identify the stained cell population and check background controls before interpreting the signal. State which source and cell population define the comparison; do not extend a cell-specific HPA result to a whole organ.

Source: UniProt O14753 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing OVOL1 staining

Immunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was used for antigen retrieval. 3, Secondary antibody was diluted at 1:200 (room temperature, 45min).

A11495-1 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172818-OVOL1/tissue/cerebral+cortex#imid_1511652

HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000172818-OVOL1/tissue/esophagus#imid_1511640

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172818-OVOL1/tissue/colon#imid_1511578

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172818-OVOL1/tissue/liver#imid_1511595

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003984; green = OVOL1, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for OVOL1 IHC & IF

🧪Run cervix first: squamous epithelial cells should stain (HPA: High in cervix squamous epithelial cells). Use adipose tissue as the negative tissue, assessing adipocytes (HPA: Not detected in adipocytes); on the cervix slide, treat adjacent nonsquamous cells as a background reference only if they remain unstained, rather than assuming they are OVOL1-negative (standard IHC practice).

Positive control tissue: Cervix (Squamous epithelial cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show OVOL1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and a host- and isotype-matched control IgG for a monoclonal antibody, or host-matched nonimmune IgG for a polyclonal antibody; confirm specificity with OVOL1 knockout material or a peptide block if the immunizing peptide is available (standard IHC practice). Quench endogenous peroxidase for chromogenic detection, and block endogenous biotin if using avidin–biotin detection (standard IHC practice).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected paraffin-section caption does not state a fixative (A11495-1 caption: fixative unreported). The caption uses Tris-EDTA at pH 9.0 for antigen retrieval, but does not establish that OVOL1 staining depends on that retrieval condition (A11495-1 caption: Tris-EDTA pH 9.0). HPA reports nucleoplasmic localization and ICC-IF images in A-431, U-251MG and U2OS, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; assess keratin-associated background in cervix squamous epithelium when interpreting staining (HPA: approved nucleoplasm localization and ICC-IF image cell lines; standard IHC/IF practice).

HPA tissue IHC evidence for OVOL1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Very low consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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
Cervix	Squamous epithelial cells	High	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	High	Protein (IHC)	HPA →
Prostate	Glandular cells	High	Protein (IHC)	HPA →
Vagina	Squamous epithelial cells	High	Protein (IHC)	HPA →
Caudate	Glial cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Appendix	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Colon	Endothelial cells	Not detected	Protein (IHC)	HPA →
Duodenum	Glandular cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced OVOL1 IHC Tips

Use the catalog antibody’s paraffin-section IHC conditions as a starting point, then judge OVOL1 staining by compartment, cell type, and controls.

What retrieval should I try when OVOL1 staining is weak in paraffin sections?

Use heat-mediated Tris-EDTA retrieval at pH 9.0 first (datasheet A11495-1). The selected paraffin-section image used that buffer, followed by primary antibody at 1:200 overnight at 4°C (caption A11495-1); use those conditions as the comparison point while adjusting heating time to your equipment. If staining remains weak, compare a citrate buffer at pH 6.0 on serial sections, keeping antibody concentration and detection identical so the retrieval change can be assessed. Evaluate nuclear signal against tissue damage and diffuse cytoplasmic staining: OVOL1 is assigned to the nucleus (UniProt O14753), while tissue IHC has uncertain reliability (HPA tissue IHC).

How should I handle fixation when OVOL1 staining varies between blocks?

OVOL1-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A11495-1). Record each block’s fixative, fixation duration, and processing history before comparing staining, because those variables can change epitope accessibility in routine IHC. Process matched control and test tissue together where possible, then hold Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 constant during troubleshooting (datasheet A11495-1; caption A11495-1). Compare nuclear staining, morphology, and background across blocks; do not assign a fixation effect to OVOL1 solely from differing tissue patterns, especially given the uncertain tissue-IHC reliability (HPA tissue IHC).

Should cytoplasmic staining count as positive OVOL1 in chromogenic IHC?

Prioritise a discrete nuclear signal: OVOL1 is assigned to the nucleus (UniProt O14753), and its approved subcellular location is nucleoplasm (HPA subcellular). Its putative transcription-factor role supports checking nuclear staining against the counterstain, while its lack of a transmembrane segment gives no basis to expect membrane staining (UniProt O14753). HPA reports cytoplasmic staining in several tissues but rates its tissue IHC uncertain because antibody staining poorly matches RNA and presumed off-target binding was observed (HPA tissue IHC). Score cytoplasmic signal separately, review the matching negative control, and require independent validation before treating that pattern as OVOL1 protein.

Could an isoform or processing difference explain inconsistent OVOL1 staining?

The supplied OVOL1 record lists no annotated isoforms, signal peptide, propeptide, glycosylation sites, or modified residues (UniProt O14753). It lists one chain spanning residues 1–267 and no transmembrane segment (UniProt O14753), so these annotations do not identify a tissue-specific processed form to explain discordant IHC. Check the antibody’s stated immunogen or epitope, if available, against the full sequence before interpreting a negative section; no epitope mapping is supplied here. Compare serial sections under the stated Tris-EDTA pH 9.0 retrieval condition (datasheet A11495-1), and use an independent antibody or orthogonal expression evidence to investigate persistent discrepancies.

How can IF help assess a questionable OVOL1 IHC pattern?

Use IF on a companion section to check whether signal overlaps nuclei, the expected OVOL1 compartment (UniProt O14753; HPA subcellular). Multiplex with a marker for the cell population being assessed; for example, a squamous epithelial marker is relevant where HPA reports high staining in squamous epithelial cells (HPA tissue IHC). Choose spectrally separated fluorophores and inspect an unstained control so tissue autofluorescence is distinguishable from specific signal. OVOL1 has no transmembrane segment and is assigned to nucleoplasm (UniProt O14753; HPA subcellular), so permeabilise sufficiently for antibody access to the nuclear epitope, while validating IF fixation and staining conditions independently of the paraffin-section IHC caption.

What controls help identify background in OVOL1 DAB staining?

Run a no-primary control through the same chromogenic workflow to reveal signal from detection reagents or tissue, including residual endogenous peroxidase activity. Include the routine peroxidase block before DAB development, and compare sections developed for the same duration so background intensity remains interpretable. Titrate around the selected image’s 1:200 primary dilution and keep retrieval at Tris-EDTA pH 9.0 while assessing whether diffuse staining falls (caption A11495-1; datasheet A11495-1). Examine edges, folds, and necrotic regions separately from intact cells; apparent cytoplasmic positivity needs particular caution because HPA reports presumed off-target binding and uncertain tissue-IHC reliability (HPA tissue IHC).

How should I quantify OVOL1 IHC across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION

Define the tissue compartment and eligible intact cells before scoring, then count nuclear staining because OVOL1 is assigned to nucleoplasm (HPA subcellular). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score calculated as the summed percentage of cells at each intensity multiplied by that intensity. Normalise cell counts to all eligible nuclei in the same compartment; if reporting positive-cell density, divide by analysed viable tissue area in mm². Keep retrieval, the selected 1:200 primary dilution, illumination, and scoring thresholds consistent across sections (datasheet A11495-1; caption A11495-1), and report cytoplasmic staining separately given HPA’s uncertain tissue-IHC assessment (HPA tissue IHC).

When should I consider an OVOL1-positive IHC field convincing?

A convincing field shows reproducible staining in intact nuclei, consistent with OVOL1’s nuclear and nucleoplasmic assignments (UniProt O14753; HPA subcellular). Check whether the stained cell population fits the tissue context: HPA reports high staining in squamous epithelial cells of cervix, esophagus, and vagina, but rates its overall tissue-IHC reliability uncertain (HPA tissue IHC). Reject staining confined to section edges, folds, or necrosis as evidence until it recurs in well-preserved tissue, and use a no-primary control to assess endogenous enzyme or detection background. Treat predominantly cytoplasmic signal cautiously because HPA reports presumed off-target binding; confirm any disputed pattern with an independent antibody or orthogonal expression evidence (HPA tissue IHC).

Boster reagents

Best OVOL1 / Putative transcription factor Ovo-like 1 IHC Antibodies

The catalog lists human, mouse, and rat IHC reactivity across its OVOL1 antibodies; A11495-1 also lists IF and has a human paraffin-section IHC figure (catalog: applications/reactivity; A11495-1 image caption).

Real IHC data Immunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was used for antigen retrieval. 3, Secondary antibody was diluted at 1:200 (room temperature, 45min).

Anti-Ovol1 Antibody

Cat # A11495-1

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A11495-1 is the SKU that will render; it lists human and mouse reactivity for IHC and IF (A11495-1 catalog: applications/reactivity). Its figure shows IHC on paraffin-embedded human colon cancer; the fixative is unreported (A11495-1 image caption).

Which to pick: For tissue IHC, choose A11495-1 when a human paraffin-section example is useful; its own figure documents that preparation, with no fixative reported (A11495-1 image caption). For IF/ICC, A11495-1 lists IF at 1:50, but supplies no IF figure or ICC-specific validation (A11495-1 catalog: applications/dilution/image alts). For rat IHC, A11495 lists rat reactivity but has no IHC figure; both catalog antibodies are rabbit polyclonals (A11495 catalog: applications/reactivity/image alts/host/dilution_raw; A11495-1 catalog: host/dilution_raw).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry O14753 (OVOL1_HUMAN, Putative transcription factor Ovo-like 1).
Human Protein Atlas. OVOL1 tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. OVOL1 subcellular location (ICC-IF): Localized to the nucleoplasm..
Human Protein Atlas. OVOL1 antibody validation summary (1 antibodies).
Evaluation of OVOL1 and Filaggrin immunohistochemical expression and clinical relevance in psoriasis. Diagnostic pathology 2024 — PMC11191297.
Ovo-Like Transcriptional Repressor 1 (OVOL1) Expression in Pleomorphic Adenoma and Carcinoma Ex Pleomorphic Adenoma: Diagnostic and Prognostic Insights. Cureus 2025 — PMC12603596.
OVOL2-Mediated ZEB1 Downregulation May Prevent Promotion of Actinic Keratosis to Cutaneous Squamous Cell Carcinoma. Journal of clinical medicine 2020 — PMC7141138.
Unveiling the impact of OVOL1 on prognosis, immune microenvironment, and proliferation in pancreatic cancer. Scientific reports 2025 — PMC12304111.
PubMed PMID:17974005 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.
PubMed PMID:9383297 — UniProt-cited evidence.