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EHMT2 / Histone-lysine N-methyltransferase EHMT2 · IHC design guide
Design Immunohistochemistry for EHMT2
Plan EHMT2 chromogenic IHC-P around the nuclear staining observed in several tissues (HPA tissue IHC). This guide identifies vaginal squamous epithelial cells as a high-staining control (HPA tissue IHC) and gives the catalog antibody’s 2–5 μg/mL IHC range (datasheet A01055-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EHMT2 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 staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining; high in vaginal squamous epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01055-2) | |
| Positive control | Vagina+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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Higher expression in fetal liver and thymus (UniProt) | |
| Isoform / epitope | 3 isoforms; verify epitope coverage across them (UniProt) |
Section 1
Recommended EHMT2 IHC & IF Protocols
The catalog antibody protocol is paired with 3 published EHMT2 IHC protocols from paraffin sections (PMC12853826; PMC13039405; PMC6254934).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01055-2) |
| Fixation | Image fixative and duration unreported (datasheet A01055-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01055-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01055-2) |
| Primary antibody | Rabbit anti-EHMT2, 2-5μg/ml (datasheet A01055-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01055-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01055-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EHMT2-positive staining in squamous epithelial cells of vagina (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A01055-2). Use each published retrieval condition with its corresponding antibody (PMC13039405; PMC6254934).
Section 2
What Is the Expected EHMT2 Staining Pattern?
EHMT2 is a nuclear, euchromatin-associated protein with no transmembrane segment (UniProt Q96KQ7: location and topology). In paraffin-section IHC, expect nuclear staining in selected epithelial populations, including strongly stained vaginal squamous cells and moderately stained colonic glandular cells (HPA: vagina High; colon Medium). HPA describes nuclear expression in several tissues, with Supported IHC reliability and medium consistency between staining and RNA data (HPA: tissue IHC).
| Distinct nuclear signal in vaginal squamous epithelium, with weaker nuclear signal in colonic glands. | This matches the reported cell types and relative staining levels (HPA: vaginal squamous cells High; colonic glandular cells Medium). Score the relevant cells and their nuclei separately from surrounding tissue; whole-section darkness can conceal the expected cell-specific pattern (general IHC practice). |
| Strong, convincing staining is mainly at cell membranes or throughout the cytoplasm. | Treat this as a compartment mismatch requiring investigation: EHMT2 is nuclear and has no transmembrane segment (UniProt Q96KQ7: location and topology; HPA: nuclear tissue expression). Review the counterstain, tissue morphology and detection controls before interpreting the signal as EHMT2 (general IHC practice). |
| Strong staining appears in adipocytes or bone-marrow hematopoietic cells while expected epithelial cells are weak. | Those populations were reported as Not detected in the supplied tissue IHC profile (HPA: adipocytes and bone-marrow hematopoietic cells Not detected). Unexpected staining can reflect cross-reactivity or endogenous detection activity; it does not by itself establish a new EHMT2 distribution (general IHC practice). |
| Brown precipitate covers stroma, empty spaces and nuclei without clear cell boundaries. | A diffuse deposit that does not track nuclei is difficult to score as EHMT2 (UniProt Q96KQ7: nuclear location; general IHC practice). Assess nonspecific binding, inadequate washing and detection background with appropriate controls before comparing tissue intensity (general IHC practice). |
| No nuclear signal appears in a well-preserved vaginal squamous epithelium section. | This conflicts with the reported High staining and merits a run-level check (HPA: vaginal squamous cells High). It is not proof that the specimen lacks EHMT2: HPA calls the tissue IHC reliability Supported, with medium staining–RNA consistency (HPA: tissue IHC); check controls and morphology first (general IHC practice). |
💡Expected EHMT2 appearanceA persuasive positive is cell-bounded nuclear chromogen in vaginal squamous cells at a relatively High level, with possible Medium nuclear staining in colonic glands; dominant membrane staining or diffuse, anatomy-independent deposit is suspect (HPA: vagina High, colon Medium and nuclear tissue expression; UniProt Q96KQ7: nuclear location; general IHC practice).
| Cell population and tissue choice | HPA reports High staining in vaginal squamous cells, Medium in several epithelial populations, Low in selected neuronal and glandular populations, and Not detected in specified others (HPA: tissue IHC). Choose and score a named cell population; a tissue-wide positive or negative label loses that distinction (general IHC practice). |
| Compartment and molecular topology | EHMT2 associates with nuclear euchromatin and has no transmembrane segment (UniProt Q96KQ7: location and topology). Nuclear localisation is therefore the primary pattern check; the supplied evidence gives no basis for treating membrane signal as the expected result (UniProt Q96KQ7: location and topology). |
| Antibody validation scope | One supplied antibody has Supported IHC status, while the other has Supported ICC status with no IHC status supplied (HPA: HPA050550 IHC Supported; HPA060259 ICC Supported). An ICC result alone does not establish performance in paraffin-section IHC (general IHC practice). |
| Retrieval and target-specific limits | Antigen retrieval is a general paraffin-section IHC variable to optimize against control tissue (general IHC practice). The supplied UniProt and HPA records give no EHMT2-specific retrieval setting or fixation-sensitivity finding; do not infer either from staining level, topology or modified residues (UniProt Q96KQ7: topology and modified residues; HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive vaginal epithelium has no nuclear signal. | The run or specimen may have failed to show the reported High signal; the cause cannot be assigned from this slide alone (HPA: vaginal squamous cells High; general IHC practice). | Verify that the expected squamous cells are present and preserved, then inspect positive and detection controls and review the paraffin IHC conditions (general IHC practice). |
| Signal is cytoplasmic or membranous instead of nuclear. | The dominant compartment conflicts with the reported nuclear localisation and nonmembrane topology (HPA: nuclear tissue expression; UniProt Q96KQ7: nucleus and no transmembrane segment). | Compare signal with the nuclear counterstain and tissue boundaries; investigate nonspecific primary or detection staining before scoring it as EHMT2 (general IHC practice). |
| Adipocytes stain strongly alongside the intended epithelium. | Strong adipocyte staining is unexpected in this HPA profile and may reflect cross-reactivity or detection background (HPA: adipocytes Not detected; general IHC practice). | Compare a reagent-appropriate negative control and the neighbouring epithelial pattern; score only convincing nuclear staining in identified cells (general IHC practice). |
| The whole section looks uniformly brown. | Widespread precipitate can obscure the reported cell-specific nuclear pattern (HPA: nuclear expression in several tissues; general IHC practice). | Inspect a negative detection control, washing and background suppression; reassess only after nuclei and cell boundaries can be distinguished (general IHC practice). |
| A lymph-node or bone-marrow section appears negative despite reported EHMT2 expression. | UniProt reports expression in examined tissues, including lymph node, and lower expression in bone marrow, while HPA reports Not detected staining in lymph-node germinal-center and bone-marrow hematopoietic cells (UniProt Q96KQ7: tissue specificity; HPA: tissue IHC). | Keep expression evidence and cell-specific IHC observations distinct; use the named HPA-positive epithelial populations to assess whether the IHC run worked (HPA: vagina High; colon Medium; general IHC practice). |
| What pattern should a separate IF/ICC result show? | HPA supports nucleoplasmic and nuclear-speckle localisation in ICC-IF images, while the tissue IHC profile describes nuclear expression (HPA: subcellular ICC-IF; HPA: tissue IHC). | Interpret nuclear fluorescence with those compartments in mind on the separate IF/ICC guide page; do not transfer an ICC-only antibody validation status to paraffin IHC (HPA: subcellular ICC-IF; HPA: HPA060259 ICC Supported, IHC status not supplied). |
Experiment evidence
IHC & IF images showing EHMT2 staining
A01055-2 · IHCIHC analysis of EHMT2/G9A using anti-EHMT2/G9A antibody (A01055-2). EHMT2/G9A was detected in a paraffin-embedded section of human liver can
A01055-2 · IHCIHC analysis of EHMT2/G9A using anti-EHMT2/G9A antibody (A01055-2). EHMT2/G9A was detected in a paraffin-embedded section of human rectal ca
A01055-2 · IFIF analysis of EHMT2/G9A using anti-EHMT2/G9A antibody (A01055-2). EHMT2/G9A was detected in an immunocytochemical section of A549 cells. En
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000204371-EHMT2/tissue/cerebral+cortex#imid_14514108
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000204371-EHMT2/tissue/colon#imid_14576720
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000204371-EHMT2/tissue/liver#imid_14576712
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000204371-EHMT2/tissue/kidney#imid_14576729
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA060259; green = EHMT2, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA060259; green = EHMT2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EHMT2 IHC & IF
🧪Run vagina first: its squamous epithelial cells should show nuclear staining (HPA: High in vaginal squamous epithelial cells; UniProt Q96KQ7: nucleus). Use adipose tissue as the negative tissue (HPA: adipocytes Not detected); on the vaginal slide, cells without EHMT2 signal should retain counterstained nuclei without brown nuclear deposit (standard IHC practice).
Positive control tissue: Vagina (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 EHMT2 in HEK293, MCF-7, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported), Nuclear speckles (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s class and concentration, and an EHMT2-knockout biological control if available (caption: rabbit primary; standard IHC practice). For the vaginal positive slide, block endogenous peroxidase and assess endogenous biotin background because the reported detection uses SABC and DAB (selected-SKU caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The reported paraffin-section staining used heat retrieval in EDTA at pH 8.0; it does not establish whether retrieval is required for every specimen (selected-SKU caption: EDTA pH 8.0). No supplied comparison establishes that frozen sections or IF are easier; in vaginal squamous epithelium, score nuclear signal and treat cytoplasmic DAB as background relative to the expected localization (HPA: High in vaginal squamous epithelial cells; UniProt Q96KQ7: nucleus; standard IHC practice).
HPA tissue IHC evidence for EHMT2
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 |
|---|---|---|---|---|
| Vagina | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced EHMT2 IHC Tips
Troubleshoot EHMT2 chromogenic IHC in paraffin sections by checking nuclear staining, tissue context and the controls behind each result (UniProt Q96KQ7; HPA tissue IHC).
How should I adjust retrieval when EHMT2 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01055-2). The catalog antibody's liver cancer tissue image used this retrieval before overnight incubation with 2 μg/ml primary antibody at 4°C (caption A01055-2). If nuclear staining is weak, vary heating time in small steps while keeping buffer and detection conditions fixed, and compare morphology with a matched section (standard IHC practice). Record the heating conditions and assess nuclear signal against a no-primary control; damaged morphology or widespread staining makes an apparent signal gain difficult to interpret (standard IHC practice; UniProt Q96KQ7 localisation).
Can fixation explain weak or uneven EHMT2 staining?
Target-specific fixation sensitivity is unknown: the selected tissue caption identifies a paraffin section but does not state its fixative (caption A01055-2). Record the fixative, fixation duration and processing history for each specimen before comparing staining across cases (standard IHC practice). Compare sections processed together using the same EDTA pH 8.0 retrieval and primary incubation, then check nuclear detail and staining uniformity (datasheet A01055-2; standard IHC practice). If an established control also stains unevenly, investigate section handling and detection before attributing the pattern to EHMT2 biology (standard IHC practice; UniProt Q96KQ7 localisation).
Should EHMT2 staining appear outside the nucleus?
Prioritise nuclear staining: EHMT2 is assigned to the nucleus and euchromatic chromosome regions (UniProt Q96KQ7), while tissue IHC reports nuclear expression in several tissues (HPA tissue IHC). Supported cell imaging places it in the nucleoplasm and nuclear speckles (HPA subcellular). Score nuclear and cytoplasmic staining separately, using the hematoxylin counterstain to define intact nuclei and excluding damaged areas (standard IHC practice). Predominantly diffuse cytoplasmic DAB staining deserves a no-primary control and a review of blocking and detection, because it differs from the documented localisation; nuclear speckles need not resolve as discrete structures in chromogenic tissue sections (UniProt Q96KQ7; HPA subcellular; standard IHC practice).
Could isoforms or epitope accessibility change the IHC pattern?
EHMT2 has 3 recorded isoforms, but the supplied antibody caption does not identify its binding epitope or establish isoform coverage (UniProt Q96KQ7; caption A01055-2). The annotated Pre-SET, SET and Post-SET regions span residues 972–1180, and multiple modified residues are recorded elsewhere in the protein (UniProt Q96KQ7). Do not infer which isoform or modification a stained nucleus contains from this antibody image alone; compare the antibody's documented immunogen with isoform sequences if that information becomes available (standard IHC practice). Keep retrieval and antibody concentration constant while checking whether a changed pattern tracks specimen processing or a separately validated antibody (standard IHC practice).
How can IF help examine EHMT2 alongside a cell-type marker?
For a separate IF assay, pair EHMT2 with a validated marker for the cell population being assessed, and inspect whether both signals occupy the expected cells (standard IF practice; UniProt Q96KQ7 localisation). Choose spectrally separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence, using single-stain controls to assess bleed-through (standard IF practice). EHMT2 has no transmembrane segment and is nuclear, so permeabilise fixed cells sufficiently to admit antibody to the nuclear epitope; 0.1% Triton X-100 for 5–10 minutes is a general starting condition to optimise (UniProt Q96KQ7; standard IF practice). Assess nucleoplasmic and nuclear-speckle patterns against a nuclear counterstain and no-primary control (HPA subcellular; standard IF practice).
What should I check when DAB obscures EHMT2 nuclei?
The selected IHC image used 10% goat serum blocking, 2 μg/ml primary antibody and biotin-based DAB detection (caption A01055-2). Compare a no-primary section to identify signal arising from the secondary, detection reagents or tissue rather than primary binding (standard IHC practice). Apply a peroxidase block, review endogenous biotin interference with the chosen detection system, and shorten chromogen development if background masks nuclei (standard IHC practice). If background persists, titrate the primary from the documented concentration while holding retrieval and exposure conditions constant; judge improvement by nuclear contrast and preserved morphology (caption A01055-2; UniProt Q96KQ7 localisation; standard IHC practice).
How should I score EHMT2 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions before scoring, and count only intact nuclei with interpretable chromogenic signal (standard IHC practice; UniProt Q96KQ7 localisation). Report the percentage of positive nuclei and an H-score based on nuclear intensity, using the same intensity thresholds across sections (standard IHC practice). Normalise counts to the total nuclei of the specified cell population; if reporting positive-cell density, divide by assessed tissue area in mm² (standard IHC practice). Exclude necrosis and folds, document sampling and staining batches, and compare cases against controls processed in the same run because the HPA profile shows cell-dependent staining levels (standard IHC practice; HPA tissue IHC).
How can I distinguish genuine EHMT2 staining from artefact?
A credible result places DAB in intact nuclei within a defined cell population, consistent with EHMT2's nuclear localisation (UniProt Q96KQ7; HPA tissue IHC). HPA reports high staining in vaginal squamous epithelial cells but no detected staining in bone marrow hematopoietic cells, illustrating why cell identity matters when selecting comparators (HPA tissue IHC). Treat staining confined to section edges, necrotic zones or predominantly cytoplasmic areas cautiously, and check a no-primary section for endogenous enzyme or detection background (standard IHC practice; UniProt Q96KQ7 localisation). Interpret differences against specimen handling and matched controls; nuclear EHMT2 staining alone does not establish its methyltransferase activity or a particular histone modification (standard IHC practice; UniProt Q96KQ7 function).
Boster reagents
Best EHMT2 / Histone-lysine N-methyltransferase EHMT2 IHC Antibodies
A01055-2 has IHC images from human liver cancer, human rectal cancer, and mouse ovary paraffin sections, plus an IF image from A549 cells (catalog image captions).
Real IHC data
Anti-EHMT2/G9A Antibody ®
Cat # A01055-2
A01055-2 is the card that will render; its IHC captions document staining in human liver cancer, human rectal cancer, and mouse ovary paraffin sections (catalog IHC image captions). Its IF caption documents staining in A549 cells, and the catalog lists human, mouse, and rat reactivity (catalog IF image caption; catalog reactivity).
Which to pick: For tissue IHC, choose A01055-2 because its own captions document paraffin-section staining at 2 μg/ml; the fixative is unreported (A01055-2 IHC image captions). For IF/ICC, A01055-2 has an A549 IF image at 5 μg/ml, while monoclonal M01055-1 lists ICC/IF without a supplied image (A01055-2 IF image caption; M01055-1 catalog applications and image list). For cross-species work, A01055-2 lists human, mouse, and rat reactivity, with IHC images for human and mouse; monoclonal M01055-1 lists the same species and IHC/IF applications but has no supplied images (catalog reactivity, applications, and image lists).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96KQ7 (EHMT2_HUMAN, Histone-lysine N-methyltransferase EHMT2).
- Human Protein Atlas. EHMT2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. EHMT2 subcellular location (ICC-IF): Localized to the nucleoplasm and nuclear speckles..
- Human Protein Atlas. EHMT2 antibody validation summary (2 antibodies).
- Down-regulation of EHMT2 through irisin-mediated epigenetic modification promotes osteogenesis via promoting DLX3 transcription. Cell communication and signaling : CCS 2026 — PMC12853826.
- EHMT2-mediated R-loop formation promotes the malignant progression of prostate cancer via activating Aurora B. Clinical and translational medicine 2025 — PMC11705492.
- The EHMT2-MBLAC2 axis suppresses ribosomal DNA transcription in response to nucleolar DNA damage. Cell death & disease 2026 — PMC13039405.
- The novel prognostic marker, EHMT2, is involved in cell proliferation via HSPD1 regulation in breast cancer. International journal of oncology 2019 — PMC6254934.
- PubMed PMID:11707778 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14656967 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EHMT2 (Histone-lysine N-methyltransferase EHMT2). Expected localisation Nuclear staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01055-2). Validated IHC antibody: A01055-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Vagina (Squamous epithelial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01055-2); 2. Fixation: Image fixative and duration unreported (datasheet A01055-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A01055-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01055-2); 8. Primary antibody: Rabbit anti-EHMT2, 2-5μg/ml (datasheet A01055-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01055-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; verify epitope coverage across them (UniProt); Regulation: Higher expression in fetal liver and thymus (UniProt). Evidence: UniProt Q96KQ7, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm and nuclear speckles., Datasheet A01055-2, PMC12853826, PMC13039405. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EHMT2/immunohistochemistry-ihc-guide.