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TET2 / Methylcytosine dioxygenase TET2 · IHC design guide
Design Immunohistochemistry for TET2
Plan TET2 staining in paraffin sections around its observed nuclear tissue pattern (HPA tissue IHC). This guide covers consistent fixation, IHC antibody selection and interpretation of staining where expression may be reduced in MDS granulocytes (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TET2 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 | Nuclei across cell types (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in cells throughout tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adipose tissue+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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00191) | |
| Caveat | Tissue staining awaits external verification (HPA tissue IHC) | |
| Regulation | Reduced in MDS granulocytes (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage unreported (UniProt; catalog datasheet) |
Section 1
Recommended TET2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published TET2 staining parameters for head and neck squamous cell carcinoma (PMC7186610) and non-functioning pituitary adenomas (PMC7381329).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A00191); 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-TET2, 2 μg/mL (datasheet A00191) |
| 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 | TET2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitously expressed in the nuclei of all cells. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min (page antigen retrieval); citrate retrieval is a published alternative (PMC7381329).
Section 2
What Is the Expected TET2 Staining Pattern?
TET2 is a nuclear protein with no transmembrane segment (UniProt Q6N021: localization, topology). In paraffin sections, expect staining in nuclei across many cell types, with high staining reported in bone marrow hematopoietic cells and several other cell populations (HPA: tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between staining and RNA expression; external verification is pending (HPA: tissue IHC reliability).
| Nuclear staining appears across multiple cell types in the section. | This fits the reported ubiquitous nuclear pattern (HPA: tissue IHC). Compare intensity within the same run: high staining is reported in bone marrow hematopoietic cells and adrenal glandular cells (HPA: High). Nuclear staining alone does not establish that every stained cell has the same TET2 abundance. |
| Strong signal lies mainly outside nuclei, with little nuclear staining. | A predominantly extranuclear IHC pattern conflicts with TET2's nuclear localization and HPA's tissue pattern (UniProt Q6N021: localization; HPA: tissue IHC). HPA also reports uncertain additional cytosolic localization by ICC-IF, so a small cytoplasmic component is less decisive than loss of the expected nuclear signal (HPA: subcellular ICC-IF). |
| Only one unexpected cell population stains strongly while neighboring nuclei are blank. | TET2 is reported as broadly expressed, and HPA describes nuclear staining in all cells (UniProt Q6N021: tissue specificity; HPA: tissue IHC). An isolated, compartmentally discordant signal warrants checks for cross-reactivity or endogenous chromogenic activity (general IHC practice). Cell-to-cell intensity differences alone do not prove a false positive. |
| A diffuse chromogenic haze obscures nuclei and tissue boundaries. | The slide cannot support a reliable TET2 localization call when background obscures the expected nuclear pattern (HPA: tissue IHC). Review the detection and blocking controls, washing, and primary antibody concentration as general IHC troubleshooting steps (general IHC practice); the supplied sources do not identify a TET2-specific cause. |
| No nuclear signal appears in bone marrow hematopoietic cells. | This differs from a reported high-staining population (HPA: High in bone marrow hematopoietic cells). Check section integrity and run controls before interpreting the absence as biological loss (general IHC practice). HPA reports medium staining/RNA consistency and pending external verification, so one negative section cannot settle TET2 status (HPA: tissue IHC reliability). |
💡Expected TET2 appearanceCall positive when clear nuclear staining is present across cell types, including strong signal in a documented high-staining population such as bone marrow hematopoietic cells (HPA: tissue IHC); diffuse haze or predominantly extranuclear chromogen is suspect (HPA: nuclear pattern; general IHC practice).
| Expected compartment | UniProt places TET2 in the nucleus and on chromosomes; chromatin localization depends on monoubiquitination at Lys-1299 (UniProt Q6N021: localization). HPA tissue IHC describes ubiquitous nuclear staining (HPA: tissue IHC). Use the nuclear compartment to judge chromogenic signal, without requiring uniform intranuclear texture. |
| Choice of reference tissue | HPA reports high staining in bone marrow hematopoietic cells, adipocytes in adipose tissue, and adrenal glandular cells, among others (HPA: High). Its supplied tissue list contains no negative or low examples (HPA: tissue IHC). Use documented high-staining populations as positive references; do not designate an unsupported tissue as TET2-negative. |
| Antibody and evidence limits | One listed antibody, HPA043135, is rated Supported for IHC (HPA: antibody validation). The tissue profile is Supported with medium staining/RNA consistency and pending external verification (HPA: tissue IHC reliability). These ratings support comparison with the reported pattern, but do not establish that every individual positive or negative cell is correctly classified. |
| Molecular forms | UniProt lists three TET2 isoforms and a single annotated 1–2002 protein chain, with no signal peptide, propeptide, or transmembrane segment (UniProt Q6N021: isoforms, processing, topology). No epitope location is supplied, so the record cannot predict isoform-specific staining or justify interpreting a compartment shift as cleavage or shedding. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclei are faint throughout the run, including a documented high-staining control. | Low overall assay sensitivity is plausible; the supplied sources do not identify a TET2-specific retrieval or fixation effect (general IHC practice; HPA: High in bone marrow hematopoietic cells). | Check control performance, antibody dilution and detection reagents, then optimize antigen retrieval within the local IHC workflow (general IHC practice). Do not assign the weak result to TET2 loss until the positive control works. |
| The chromogen is mainly cytoplasmic. | This conflicts with nuclear tissue IHC, although additional cytosolic localization in HPA ICC-IF is marked uncertain (HPA: tissue IHC; HPA: subcellular ICC-IF). | Check the nuclear counterstain and compare compartments in a documented high-staining tissue (general IHC practice; HPA: High in bone marrow hematopoietic cells). Treat persistent cytoplasmic-only IHC as unresolved specificity. |
| Background is widespread even where cellular detail is poor. | Nonspecific antibody binding or detection background can obscure compartment scoring (general IHC practice). The supplied TET2 sources do not specify which mechanism applies. | Inspect a no-primary control, blocking and wash conditions, and the detection system; adjust primary antibody concentration using the local assay's controls (general IHC practice). |
| Signal remains in a no-primary control. | Primary-independent detection or endogenous chromogenic activity is possible (general IHC practice). This control cannot identify TET2 expression. | Troubleshoot the detection chemistry and appropriate endogenous-activity blocking before scoring the TET2 slide (general IHC practice). |
| One cell type appears negative while adjacent cells stain. | True intensity variation or technical variation may contribute; HPA describes ubiquitous nuclear staining but reports only medium staining/RNA consistency (HPA: tissue IHC). | Confirm nuclear morphology, compare same-run positive controls, and repeat or corroborate a consequential cell-type call (general IHC practice). Avoid claiming a validated negative cell population from this payload. |
| The observed pattern differs from HPA images despite an acceptable control. | HPA's tissue interpretation is Supported but pending external verification, and the supplied antibody list gives only one IHC Supported entry (HPA: tissue IHC reliability; HPA: antibody validation). | Record the discrepancy by cell type and compartment, review assay controls, and seek independent corroboration before assigning biological meaning (general IHC practice). |
Experiment evidence
IHC & IF images showing TET2 staining
A00191 · IHCImmunohistochemistry of TET2 in human colon carcinoma tissue with TET2 antibody at 2 μg/ml.
A00191 · IFImmunofluorescence of TET2 in human colon carcinoma tissue with TET2 antibody at 20 μg/ml. Green: TET2 Antibody (A00191) Blue: DAPI staining
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168769-TET2/tissue/cerebral+cortex#imid_11539923
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168769-TET2/tissue/colon#imid_11539849
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168769-TET2/tissue/liver#imid_11539850
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000168769-TET2/tissue/kidney#imid_11539851
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043135; green = TET2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043135; green = TET2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TET2 IHC & IF
🧪Run bone marrow first: hematopoietic cells should show TET2 staining (HPA: High in bone marrow hematopoietic cells). HPA detects TET2 in all 45 scored tissues, so use no-primary and isotype controls in place of a negative tissue; anucleate erythrocytes on the positive slide should lack nuclear staining (HPA: no negative tissue; standard IHC practice: erythrocytes lack nuclei).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: TET2 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 ICC-IF images show TET2 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and monoclonal or polyclonal format, and matched TET2-knockout material as a biological negative (standard IHC practice). For bone marrow chromogenic IHC, quench endogenous peroxidase and block endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A00191 colon carcinoma caption does not state a fixative (selected-SKU IHC caption). Retrieval dependence is unreported; compare standard antigen-retrieval conditions on paraffin sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; IF images support nucleoplasmic localization, while cytosolic localization is uncertain, and marrow requires attention to endogenous peroxidase background (HPA: subcellular; standard IHC practice).
HPA tissue IHC evidence for TET2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: TET2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TET2 IHC Tips
Troubleshoot TET2 staining in paraffin sections by checking retrieval, nuclear localisation, background controls and cell-based scoring before interpreting chromogenic signal.
How should I retrieve TET2 when nuclear staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting: nuclear antigen). Let sections cool consistently, then compare staining with a known positive section from the same run; TET2 is expected in nuclei and on chromosomes (UniProt Q6N021 localisation). If nuclear signal remains weak, test a shorter or longer heating interval on adjacent sections while keeping the antibody concentration and detection system fixed (general IHC optimisation practice). Record nuclear signal and tissue integrity together, because damaged morphology makes compartment-based interpretation unreliable (general IHC interpretation practice).
Could fixation explain weak or uneven TET2 staining?
TET2-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence (selected A00191 caption: fixative not stated). Compare sections with documented processing histories and hold retrieval, antibody concentration and detection conditions constant while assessing nuclear signal (general IHC optimisation practice). The selected antibody image shows TET2 staining in human colon carcinoma tissue at 2 µg/ml, but its caption does not identify the fixative (selected A00191 caption). If signal varies across blocks, record fixation details where available and assess morphology and edge effects before attributing the difference to TET2 abundance (general IHC interpretation practice).
What should I check when TET2 appears cytoplasmic?
Score nuclear staining first: TET2 is annotated in the nucleus and on chromosomes, with chromatin localisation dependent on monoubiquitination at Lys-1299 (UniProt Q6N021 localisation). A supported nucleoplasmic location and an uncertain additional cytosolic location are reported, so isolated cytoplasmic colour needs cautious interpretation (HPA subcellular). Compare the nuclear and cytoplasmic patterns against a matched negative control and inspect whether colour follows damaged tissue or nonspecific deposits (general IHC interpretation practice). TET2 has no annotated transmembrane segment, so a crisp membrane-only pattern would prompt a specificity and detection review (UniProt Q6N021 topology; general IHC interpretation practice).
How can isoforms or epitope accessibility affect TET2 IHC?
Check the antibody's documented immunogen or epitope before interpreting discordant staining, because the supplied record lists 3 TET2 isoforms without mapping this antibody's binding site (UniProt Q6N021 isoforms; supplied antibody evidence). TET2 is a 2002-amino-acid protein with annotated phosphorylation sites, including Ser-15 and Ser-1107 (UniProt Q6N021 sequence and modified residues). Those annotations alone do not establish that a particular isoform or modification changes staining with this antibody (supplied antibody evidence). Compare retrieval conditions on adjacent sections and seek antibody-specific epitope information before calling a staining difference isoform-specific (general IHC validation practice).
How should I use IF to investigate an ambiguous IHC pattern?
Use IF as a companion localisation check for ambiguous chromogenic TET2 staining, while evaluating the IHC section on its own controls (general IHC/IF practice). Multiplex TET2 with a validated marker for the expected cell population, such as a granulocyte marker when examining hematopoietic material; granulocytes have the highest reported TET2 expression (UniProt Q6N021 tissue specificity). Choose fluorophores in channels with low specimen autofluorescence and include single-colour and no-primary controls (general IF practice). Because TET2 is primarily nuclear and has no transmembrane segment, optimise permeabilisation for intracellular access without assuming a membrane-facing epitope (UniProt Q6N021 localisation and topology; general IF practice).
How do I reduce diffuse brown staining without losing TET2 signal?
First inspect a no-primary control and verify that peroxidase blocking and washes are effective before changing the TET2 antibody conditions (general chromogenic IHC practice). Assess whether diffuse DAB colour appears outside nuclei, since widespread nuclear staining is reported for TET2 in tissue (HPA tissue IHC profile). For SKU A00191, 2 µg/ml is the concentration stated for the human colon carcinoma image, rather than a universal working concentration (selected A00191 caption). If background persists, compare a modest concentration reduction and blocking conditions on adjacent sections, then retain the condition that preserves interpretable nuclear staining (general IHC optimisation practice).
How should I quantify TET2 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then measure the percentage of TET2-positive nuclei and nuclear intensity using a consistent threshold (general IHC quantification practice; UniProt Q6N021 localisation). An H-score can combine percentage and intensity, while positive nuclei per mm² can describe spatial density when cell counts differ (general IHC quantification practice). Normalise positive counts to the total evaluable nuclei in the same region, and report analysed area when using density (general IHC quantification practice). Keep retrieval, detection, exposure to chromogen and scoring rules consistent across compared sections (general IHC quantification practice).
Which patterns support genuine TET2 staining rather than artefact?
A reproducible nuclear pattern in intact cells is consistent with TET2's annotated nuclear localisation and the reported broad nuclear tissue profile (UniProt Q6N021 localisation; HPA tissue IHC profile). Strong signal in hematopoietic cells can also fit the reported expression pattern, particularly in granulocytes (UniProt Q6N021 tissue specificity). Treat membrane-only colour, staining concentrated at cut edges or necrotic areas, and colour reproduced in no-primary controls as reasons to investigate artefact or endogenous enzyme activity (UniProt Q6N021 topology; general chromogenic IHC practice). The HPA tissue assessment is Supported but pending external verification, so confirm consequential interpretations with appropriate controls and independent evidence (HPA tissue IHC reliability).
Boster reagents
Best TET2 / Methylcytosine dioxygenase TET2 IHC Antibodies
A00191 has IHC and IF images of human colon carcinoma tissue (catalog: A00191 image captions). Its listed reactivity is human (catalog: A00191 reactivity).
Real IHC data
Anti-TET2 Antibody
Cat # A00191
A00191 is the SKU with a rendered card and is listed for human IHC-P and IF (catalog: A00191 applications and reactivity). Its own captions show IHC and IF staining in human colon carcinoma tissue (catalog: A00191 image captions).
Which to pick: For tissue IHC, choose A00191: its IHC caption shows human colon carcinoma tissue at 2 μg/mL; the fixative is unreported (catalog: A00191 IHC image caption). For IF, A00191 has an image at 20 μg/mL in human colon carcinoma tissue; for ICC, A00191-2 lists ICC/IF but has no image caption (catalog: A00191 IF image caption; A00191-2 applications and image captions). For mouse or rat samples, A00191-2 lists both species with IHC and ICC/IF, while A00191-3 lists mouse with IHC; clonality is unreported for both (catalog: A00191-2 and A00191-3 reactivity, applications and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6N021 (TET2_HUMAN, Methylcytosine dioxygenase TET2).
- Human Protein Atlas. TET2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. TET2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. TET2 antibody validation summary (1 antibodies).
- Restoration of TET2 deficiency inhibits tumor growth in head neck squamous cell carcinoma. Annals of translational medicine 2020 — PMC7186610.
- Changes in DNA 5-Hydroxymethylcytosine Levels and the Underlying Mechanism in Non-functioning Pituitary Adenomas. Frontiers in endocrinology 2020 — PMC7381329.
- Short- and long-term effects of (56)Fe irradiation on cognition and hippocampal DNA methylation and gene expression. BMC genomics 2016 — PMC5078898.
- Bi-directional and shared epigenomic signatures following proton and (56)Fe irradiation. Scientific reports 2017 — PMC5579159.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TET2 (Methylcytosine dioxygenase TET2). Expected localisation Nuclei across cell types (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A00191 at 2 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. 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 A00191); 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-TET2, 2 μg/mL (datasheet A00191); 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 A00191); Caveat: Tissue staining awaits external verification (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage unreported (UniProt; catalog datasheet); Regulation: Reduced in MDS granulocytes (UniProt). Evidence: UniProt Q6N021, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A00191, PMC7186610, PMC7381329. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TET2/immunohistochemistry-ihc-guide.