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TET3 / Methylcytosine dioxygenase TET3 · IHC design guide
Design Immunohistochemistry for TET3
Plan TET3 IHC-P around cytoplasmic tissue staining (HPA tissue IHC) and its annotated nuclear localisation (UniProt). Start the catalog antibody at 2.5 μg/mL (datasheet), using esophageal squamous epithelial cells as a high-staining reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TET3 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 in tissue (HPA tissue IHC); nuclear by annotation (UniProt) | |
| Staining pattern | Strong cytoplasmic staining in squamous epithelial and smooth muscle cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Esophagus+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 A03371) | |
| Caveat | Tissue staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Tissue-enhanced RNA: bone marrow, skin (HPA RNA) | |
| Isoform / epitope | 3 isoforms (UniProt); epitope coverage unspecified (datasheet) |
Section 1
Recommended TET3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published TET3 IHC methods for human ovarian tissue microarrays and tumor sections (PMC4855705; PMC13169958).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03371); 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-TET3, 2.5 μg/mL (datasheet A03371) |
| 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 | TET3-positive staining in squamous epithelial cells of esophagus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several different tissue types, highly abundant in squamous epithelial and smooth muscle cells. No signal in the no-primary control. |
💡Decision noteStart with this page’s Tris-EDTA pH 9.0 retrieval for nuclear TET3 (page retrieval; UniProt O43151). Citrate pH 6.0 is a published alternative (PMC4855705; PMC13169958).
Section 2
What Is the Expected TET3 Staining Pattern?
TET3 is annotated in the nucleus, cytoplasm and chromosomes, with no transmembrane segment (UniProt O43151). In tissue IHC, expect mainly cytoplasmic staining in selected cells, especially squamous epithelial and smooth muscle cells (HPA: tissue IHC profile). Treat that pattern as provisional: HPA rates the tissue IHC result Uncertain because antibody staining has low consistency with RNA expression (HPA: tissue IHC reliability).
| Clear cytoplasmic stain in esophageal or oral squamous epithelial cells, with nearby structures remaining distinct. | This fits a documented high staining pattern (HPA: esophagus and oral mucosa, High). Record the stained cell population and compartment alongside intensity; HPA's overall tissue IHC reliability remains Uncertain (HPA: tissue IHC reliability). |
| Nuclear staining accompanies, or exceeds, cytoplasmic staining in an otherwise plausible cell population. | Nuclear TET3 is biologically plausible (UniProt O43151: nucleus and chromosome; HPA: ICC-IF nucleoplasm, approved). It differs from HPA's mainly cytoplasmic tissue IHC profile, so assess antibody controls before treating the nuclear IHC pattern as confirmed (HPA: tissue IHC profile). |
| Signal outlines cell membranes or appears extracellular, without a convincing intracellular pattern. | That compartment does not match the annotated nuclear or cytoplasmic locations (UniProt O43151; HPA: tissue IHC profile). Consider nonspecific staining or detection artefact; TET3 has no annotated transmembrane segment or signal peptide (UniProt O43151 topology and processing). |
| Strong stain appears in adipocytes or liver cholangiocytes. | HPA reports those cell types as Not detected (HPA: adipose tissue adipocytes; liver cholangiocytes). Check for cross-reactivity or endogenous detection activity before calling the signal TET3; an HPA negative is a reference observation, not proof that every specimen must be negative (HPA: tissue IHC reliability Uncertain). |
| Esophageal squamous epithelium has no detectable stain. | That conflicts with HPA's High observation for this cell population (HPA: esophagus squamous epithelial cells). First assess section quality, retrieval and reagent performance using general IHC controls. A failed control slide cannot establish that the test specimen lacks TET3 (general IHC practice). |
💡Expected TET3 appearanceA persuasive IHC positive is distinct cytoplasmic staining in squamous epithelial cells, potentially strong in esophagus or oral mucosa (HPA: tissue IHC profile; esophagus and oral mucosa, High); membrane-only, extracellular or widespread cell-independent color calls for artefact checks (UniProt O43151 localization; general IHC practice).
| Cell population and compartment | Compare each compartment within a named cell type: HPA describes abundant cytoplasmic staining in squamous epithelial and smooth muscle cells, while its listed High examples include skin keratinocytes and placental trophoblastic cells (HPA: tissue IHC profile; skin and placenta, High). |
| Strength of the tissue reference | HPA rates tissue IHC reliability Uncertain because staining and RNA expression show low consistency (HPA: tissue IHC reliability). Its single listed antibody, HPA050845, is Uncertain for IHC and Approved for ICC; these are application-specific assessments (HPA: antibody validation). |
| Topology and processing | The annotated TET3 chain spans residues 1–1795; no signal peptide, propeptide or transmembrane segment is annotated (UniProt O43151 topology and processing). These annotations support checking unexpected surface or extracellular signal, but do not identify the antibody's epitope. |
| Isoforms | UniProt lists 3 isoforms (UniProt O43151: isoforms). Without an epitope or isoform-specific antibody record here, a staining difference cannot be assigned to a particular isoform; compare like tissues and controls before making that interpretation. |
| IF/ICC: What pattern should I expect? | HPA reports mainly nucleoplasmic signal, with additional vesicle and cytosol signal, in ICC-IF (HPA: subcellular location, approved). That supports an IF localization check but does not override the mainly cytoplasmic tissue IHC profile (HPA: tissue IHC profile, Uncertain). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control is blank or very weak. | The expected High esophageal squamous-cell signal is absent (HPA: esophagus, High); an IHC workflow failure is possible (general IHC practice). | Review section integrity, retrieval conditions, antibody dilution and detection controls using the antibody's IHC-P instructions (general IHC practice). Reassess the specimen only after a control stains as expected. |
| All tissue compartments show diffuse brown color. | Widespread color without cell boundaries does not resemble HPA's cell-selective cytoplasmic profile (HPA: tissue IHC profile); background or nonspecific detection is possible (general IHC practice). | Compare a no-primary control, inspect blocking and washes, and check the chromogen development time (general IHC practice). Score only resolved cellular stain against the counterstain. |
| Membrane rims dominate the positive cells. | A membrane-only pattern conflicts with the annotated nuclear and cytoplasmic locations and lack of a transmembrane segment (UniProt O43151). | Inspect the same cell types on the no-primary control and review edge staining or precipitate under the microscope (general IHC practice). Do not count membrane rims as a TET3 positive without independent support. |
| Cells listed as Not detected stain strongly. | Strong signal in adipocytes or cholangiocytes disagrees with HPA's tissue observations (HPA: adipose tissue; liver). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Run a no-primary control and, for peroxidase detection, check endogenous peroxidase blocking (general IHC practice). Because HPA rates tissue IHC Uncertain, report the discrepancy rather than automatically rejecting the specimen (HPA: tissue IHC reliability). |
| Nuclear staining is prominent while cytoplasmic staining is faint. | Nuclear localization has support in UniProt and ICC-IF, whereas HPA's tissue IHC summary emphasizes cytoplasm (UniProt O43151 localization; HPA: ICC-IF subcellular and tissue IHC profile). | Document nuclear and cytoplasmic scores separately, review controls, and avoid assigning a nuclear IHC pattern the same confidence as HPA's approved ICC-IF result (HPA: antibody validation and subcellular location). |
| A low-staining tissue is used as the only positive control. | HPA lists only Low staining in adrenal gland glandular cells and lung alveolar cells, for example (HPA: adrenal gland; lung). Weak or absent color there gives a limited check of assay performance. | Include a documented High cell population, such as skin keratinocytes or esophageal squamous epithelial cells, on a control section (HPA: skin and esophagus, High). Judge the assay from the stained cell type, not the tissue label alone. |
Experiment evidence
IHC & IF images showing TET3 staining
A03371 · IHCImmunohistochemistry of TET3 in human brain tissue with TET3 antibody at 2.5 μg/ml.
A03371 · IFImmunofluorescence of TET3 in human brain tissue with TET3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000187605-TET3/tissue/cerebral+cortex#imid_14260012
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000187605-TET3/tissue/colon#imid_14259922
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000187605-TET3/tissue/liver#imid_14259923
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000187605-TET3/tissue/kidney#imid_14259932
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA050845; green = TET3, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA050845; green = TET3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TET3 IHC & IF
🧪Run esophagus first and confirm staining in squamous epithelial cells (HPA: High); use adipose tissue adipocytes as the negative tissue (HPA: Not detected). On the positive slide, cells outside the scored squamous epithelium should show only background in the control stains; their TET3-negative status is unreported (HPA: esophageal squamous epithelial cells High).
Positive control tissue: Esophagus (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 TET3 in MCF-7, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and TET3 knockout material as a biological negative (standard IHC practice). For chromogenic esophagus IHC, quench endogenous peroxidase and check the control slide for residual tissue background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval requirement are unreported in the supplied evidence; assess retrieval empirically (supplied target/application evidence). The selected A03371 brain IHC caption reports 2.5 μg/ml but does not report a fixative (selected-SKU tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC, or identify an esophagus-specific artefact; IF images show mainly nucleoplasmic TET3 with additional vesicular and cytosolic localization (HPA subcellular).
HPA tissue IHC evidence for TET3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Skin | Keratinocytes | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TET3 IHC Tips
Troubleshoot TET3 staining in paraffin sections by checking retrieval, cellular compartment, controls and scoring before interpreting chromogenic signal.
What retrieval should I use when TET3 staining is weak?
Start with Tris-EDTA, pH 9.0, heat-induced antigen retrieval at 95–98 °C for 20 minutes (page retrieval setting). If staining remains weak, compare a milder citrate buffer, pH 6.0, on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Include a section known to stain in the same run, and assess nuclear and cytoplasmic signal separately because both locations are reported for TET3 (UniProt O43151 subcellular location; HPA subcellular). Excessive retrieval can damage morphology or increase diffuse background, so judge any gain in signal against tissue integrity and the matched negative control (standard IHC practice).
Could fixation explain inconsistent TET3 staining across paraffin blocks?
Target-specific fixation sensitivity for TET3 is unknown from the supplied evidence; do not infer it from tissue staining patterns or protein annotations (supplied evidence). Record the fixative and fixation duration for each block, then compare sections processed with the same retrieval and detection settings (standard IHC practice). The catalog image shows TET3 staining in human brain at 2.5 µg/ml, but its caption does not state the fixative (catalog antibody A03371 caption). If blocks differ, first assess preserved morphology and control staining before assigning a weak result to biology; parallel processing can help isolate a preparation effect (standard IHC practice).
Should I score nuclear or cytoplasmic TET3 staining?
Score nuclear and cytoplasmic staining separately, with an explicit rule for each compartment before reviewing the slides (standard IHC practice). TET3 is annotated in the nucleus, cytoplasm and chromosome, while cell imaging places it mainly in the nucleoplasm with additional cytosolic and vesicular signal (UniProt O43151 subcellular location; HPA subcellular). HPA tissue IHC describes cytoplasmic expression across several tissue types, but rates its staining reliability uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC). Use a nuclear counterstain and inspect intact cells at high magnification so overlapping cytoplasm or deposited chromogen is not mistaken for nuclear signal (standard IHC practice).
How should I handle an unknown TET3 epitope and multiple isoforms?
Treat isoform coverage as unresolved until the antibody epitope or immunogen sequence is available: the record lists 3 TET3 isoforms but supplies no antibody epitope map (UniProt O43151 isoforms; supplied antibody evidence). Check whether the mapped sequence is shared across the isoforms before describing a stain as total TET3 (standard antibody validation practice). TET3 has no annotated transmembrane segment or glycosylation sites, which does not establish how fixation or retrieval exposes this particular epitope (UniProt O43151 topology and glycosylation). If interpretation depends on an isoform, compare an independently validated isoform-specific reagent or an orthogonal assay using matched material (standard validation practice).
How can I check TET3 localisation by multiplex IF?
On the separate IF/ICC workflow, pair TET3 with a validated marker for the cell population being examined and a DNA counterstain to distinguish nucleoplasm from neighboring cytoplasm (standard IF practice; HPA subcellular). Choose spectrally separated fluorophores after inspecting unstained tissue, assigning a brighter, longer-wavelength channel to a weak target when autofluorescence is troublesome (standard IF practice). Use a mild, titrated permeabilisation step for access to nuclear TET3, then verify that cell borders and nuclear morphology remain intact; TET3 has no transmembrane segment (standard IF practice; UniProt O43151 topology). Include single-stain and secondary-only controls to check bleed-through and nonspecific fluorescence before comparing the IF pattern with chromogenic IHC (standard IF practice).
What causes diffuse brown signal in a TET3 IHC section?
Check a no-primary control first to identify staining from detection reagents, endogenous peroxidase or pigment before changing the TET3 antibody concentration (standard chromogenic IHC practice). For a peroxidase and DAB workflow, apply the routine peroxidase block, titrate primary antibody and detection reagent, and compare washes on adjacent sections (standard chromogenic IHC practice). The catalog caption reports staining at 2.5 µg/ml in human brain, but does not establish an optimal concentration for every paraffin preparation (catalog antibody A03371 caption). Distinguish granular or edge-localized deposit from signal contained within intact nuclei or cytoplasm, both reported TET3 locations (standard IHC practice; UniProt O43151 subcellular location).
What is a defensible way to quantify TET3 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear and cytoplasmic compartments independently before quantification, because both locations are reported for TET3 (standard IHC practice; UniProt O43151 subcellular location). For nuclear staining, report the percentage of positive intact cells and, where intensity is reproducible, an H-score from 0–300 using 0–3 intensity bins (standard IHC scoring practice). For spatial comparisons, report positive cells per mm² of viable tissue and normalize counts to the number of eligible cells or analyzed viable area (standard image-analysis practice). Keep thresholds, exposure or scan settings, tissue exclusions and control criteria fixed across groups; HPA rates its tissue IHC reliability uncertain (standard IHC practice; HPA tissue IHC).
How can I distinguish genuine TET3 staining from an artefact?
Favor signal within intact nuclei or cytoplasm over staining confined to section edges, folds or necrotic areas; nucleus and cytoplasm are documented TET3 locations (UniProt O43151 subcellular location; standard IHC practice). Check the no-primary control for endogenous enzyme activity or pigment, and require plausible cellular boundaries before calling a DAB-positive cell (standard chromogenic IHC practice). A strongly stained squamous epithelial population is compatible with HPA tissue observations, but that source labels TET3 tissue IHC reliability uncertain because staining and RNA data agree poorly (HPA tissue IHC). Treat unexpected compartment or cell-type patterns as provisional until replicated with a validated control or an independent assay (standard antibody validation practice).
Boster reagents
Best TET3 / Methylcytosine dioxygenase TET3 IHC Antibodies
A03371 has IHC and IF images from human brain tissue (catalog image captions) and lists human, mouse, and rat reactivity (catalog reactivity).
Real IHC data
Anti-TET3 Antibody
Cat # A03371
A03371 is listed for IHC-P and IF (catalog applications). Its images show IHC at 2.5 μg/mL and IF at 20 μg/mL, both in human brain tissue (A03371 image captions).
Which to pick: Choose A03371 for paraffin-section IHC (catalog applications: IHC-P); its own IHC image shows human brain tissue, but the fixative is unreported (A03371 IHC image caption). For IF, choose A03371 (catalog applications: IF); ICC validation is not listed (catalog applications). For cross-species studies, A03371 lists human, mouse, and rat reactivity (catalog reactivity), though its images show human tissue only (A03371 image captions); clonality is unreported.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43151 (TET3_HUMAN, Methylcytosine dioxygenase TET3).
- Human Protein Atlas. TET3 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TET3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to vesicles and cytosol..
- Human Protein Atlas. TET3 antibody validation summary (1 antibodies).
- ncRNAs-mediated overexpression of TET3 predicts unfavorable prognosis and correlates with immunotherapy efficacy in breast cancer. Heliyon 2024 — PMC10838756.
- TET3 inhibits TGF-β1-induced epithelial-mesenchymal transition by demethylating miR-30d precursor gene in ovarian cancer cells. Journal of experimental & clinical cancer research : CR 2016 — PMC4855705.
- Berberine inhibits the Warburg effect through TET3/miR-145/HK2 pathways in ovarian cancer cells. Journal of Cancer 2021 — PMC7738813.
- Dual-targeted lipid nanoparticles for TET3 siRNA delivery: nanobiotechnology strategy to remodel tumor immune microenvironment in hepatocellular carcinoma. Journal of nanobiotechnology 2026 — PMC13169958.
- PubMed PMID:23217707 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TET3 (Methylcytosine dioxygenase TET3). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); nuclear by annotation (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A03371 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Esophagus (Squamous epithelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A03371); 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-TET3, 2.5 μg/mL (datasheet A03371); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03371); Caveat: Tissue staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms (UniProt); epitope coverage unspecified (datasheet); Regulation: Tissue-enhanced RNA: bone marrow, skin (HPA RNA). Evidence: UniProt O43151, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to…, Datasheet A03371, PMC4855705, PMC13169958. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TET3/immunohistochemistry-ihc-guide.