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EHMT2 / Histone-lysine N-methyltransferase EHMT2 · IHC design guide
Design Immunohistochemistry for EHMT2
Plan chromogenic EHMT2 IHC in paraffin sections around nuclear staining and a vaginal squamous epithelial positive control (HPA tissue IHC). Use the IHC-validated antibody and score nuclear signal by cell type, with consistent fixation and appropriate controls (standard IHC practice).
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 | Strong nuclear stain in vaginal squamous 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Some cell populations have no detectable stain (HPA tissue IHC) | |
| Regulation | High in fetal liver and thymus (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended EHMT2 IHC & IF Protocols
The catalog antibody protocol is paired with three published EHMT2 IHC protocols covering mouse femur, paraffin-embedded tissue, and human breast tumors (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 EDTA pH 8.0 heat retrieval (datasheet A01055-2). If staining is weak, compare the published citrate and pH 9 retrieval conditions (PMC13039405; PMC6254934).
Section 2
What Is the Expected EHMT2 Staining Pattern?
EHMT2 is a nuclear, chromosome-associated protein with no transmembrane segment (UniProt Q96KQ7: subcellular location and topology). In paraffin-section IHC, expect nuclear staining in selected epithelial cells, including high staining in vaginal squamous epithelium and medium staining in colonic glandular cells (HPA: tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between staining and RNA data (HPA: tissue IHC reliability).
| Distinct nuclear stain in vaginal squamous epithelial cells; weaker nuclear stain in colonic glands. | This matches HPA's High rating for vaginal squamous cells and Medium rating for colonic glandular cells (HPA: tissue IHC). Compare nuclei within the relevant cell population; these ratings describe observed patterns, not required intensities for every section. |
| Predominantly cytoplasmic or membrane-like stain, with little nuclear signal. | This conflicts with the expected nuclear location (UniProt Q96KQ7: subcellular location; HPA: nuclear tissue expression). Treat it as suspect until nuclear staining is demonstrated in an HPA-positive cell population. Check the detection and counterstain in parallel (general IHC practice). |
| Strong stain in a cell population HPA lists as Not detected, such as adipocytes. | Consider cross-reactivity or endogenous detection activity, especially if the signal lacks nuclear restriction (HPA: adipocytes Not detected; UniProt Q96KQ7: nuclear location). HPA's rating is an observed result, not proof that every adipocyte lacks EHMT2; inspect appropriate detection controls (general IHC practice). |
| Diffuse stain across nuclei, cytoplasm and extracellular spaces. | Diffuse background obscures the compartment needed to interpret EHMT2 IHC (UniProt Q96KQ7: nuclear location). Review antibody dilution, blocking and wash conditions against the assay controls; nonspecific binding or detection background are possibilities (general IHC practice). |
| No nuclear signal in vaginal squamous epithelium. | That conflicts with HPA's High staining observation in those cells (HPA: vagina tissue IHC). Check that the intended epithelium is present and review antibody, retrieval and detection conditions (general IHC practice). One negative section alone cannot establish biological absence. |
💡Expected EHMT2 appearanceCall a result consistent with EHMT2 when staining is predominantly nuclear in the relevant epithelial cells, potentially High in vaginal squamous cells or Medium in colonic glands (HPA: tissue IHC; UniProt Q96KQ7: nuclear location); dominant cytoplasmic, membrane-like or diffuse stain is suspect.
| Cell type and tissue | HPA reports High staining in vaginal squamous cells, Medium in several glandular and respiratory epithelial populations, and Not detected in selected populations including adipocytes (HPA: tissue IHC). Score the named cell population rather than assigning one intensity to an entire organ. |
| Nuclear distribution | UniProt places EHMT2 in the nucleus and associates it with euchromatic chromosome regions (UniProt Q96KQ7: subcellular location). HPA describes nuclear expression across several tissues (HPA: tissue IHC); its ICC-IF data support nucleoplasm and nuclear speckles (HPA: subcellular ICC-IF). |
| Antibody evidence | The listed IHC antibody HPA050550 has Supported IHC status; HPA060259 has Supported ICC status, with no IHC status supplied for it (HPA: antibody validation). HPA describes overall tissue IHC reliability as Supported with medium staining–RNA consistency (HPA: tissue IHC reliability). |
| Isoforms and processing | UniProt lists three EHMT2 isoforms, no signal peptide or propeptide, and no transmembrane segment (UniProt Q96KQ7: isoforms, processing and topology). The payload gives no antibody epitope, so isoform recognition cannot be predicted; do not infer shedding or a fixation effect. |
| IF/ICC Q: Where should EHMT2 appear? | A: HPA supports nucleoplasmic and nuclear-speckle localisation in ICC-IF (HPA: subcellular ICC-IF). The listed ICC-supported antibody is HPA060259 (HPA: antibody validation). Those cell-image findings inform localisation; they do not specify an IF protocol for this IHC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| The suspected positive epithelium looks unstained. | The section may lack the intended cells, or assay conditions may have failed; HPA reports High staining specifically in vaginal squamous cells (HPA: tissue IHC). | Identify the cell layer on the counterstain, then check an HPA-positive tissue alongside the sample. Review retrieval, antibody dilution and detection against the chosen IHC protocol (general IHC practice); no EHMT2-specific retrieval setting is supplied. |
| All compartments look brown. | General background can hide nuclear localisation (UniProt Q96KQ7: nuclear location). Excess antibody signal or detection background is possible (general IHC practice). | Compare with an appropriate detection control, then review blocking, dilution and washes (general IHC practice). Interpret EHMT2 only where a nuclear pattern can be resolved (UniProt Q96KQ7: nuclear location). |
| Cytoplasm stains more strongly than nuclei. | The distribution conflicts with UniProt's nuclear assignment and HPA's nuclear tissue profile (UniProt Q96KQ7: subcellular location; HPA: tissue IHC). The image alone cannot identify the artefact. | Recheck cell boundaries and counterstain; compare an HPA-positive tissue and a detection control under the same staining run (general IHC practice). |
| A listed Not detected cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA lists adipocytes as Not detected but rates tissue consistency only Medium (HPA: tissue IHC). | Confirm that the signal is nuclear, examine detection controls and compare an HPA-positive cell population. Treat the discrepancy as unresolved rather than declaring the HPA-negative population biologically impossible. |
| Weak staining is seen in one tissue but strong staining in another. | HPA reports cell-specific differences, including High vaginal squamous and Medium colonic glandular staining (HPA: tissue IHC). Intensity alone does not identify assay failure. | Score each named cell population and nuclear compartment separately; compare sections stained in the same run (HPA: tissue IHC; UniProt Q96KQ7: nuclear location; general IHC practice). |
| Nuclear signal appears as speckles in an IF/ICC image. | Nucleoplasm and nuclear speckles are supported ICC-IF locations (HPA: subcellular ICC-IF). This imaging pattern does not define chromogenic IHC intensity. | For IHC, assess nuclear staining in the relevant tissue cells (HPA: tissue IHC). Use the separate IF/ICC guide for that application; this section provides no IF protocol. |
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: squamous epithelial cells should show EHMT2 staining (HPA: High). Use adipose tissue as the negative comparator, where adipocytes are not detected (HPA: Not detected); on the positive slide, cells without nuclear signal provide internal background reference, but the supplied evidence does not identify a specific negative cell type.
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 concentration-matched rabbit IgG isotype control, and EHMT2-knockout tissue or a validated peptide-block control (caption: rabbit primary antibody; standard IHC controls). For chromogenic detection, quench endogenous peroxidase and check for endogenous biotin background because the reported method uses biotinylated secondary antibody, streptavidin–biotin detection, and DAB (selected-SKU caption).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative used for the selected-SKU paraffin section is unreported (selected-SKU caption). The reported IHC method uses heat-mediated retrieval in EDTA at pH 8.0; its dependence on retrieval has not been established by a comparison (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF are easier, or document a vagina-specific artefact; HPA does report nucleoplasmic and nuclear-speckle localisation in ICC-IF images (HPA subcellular).
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 staining in paraffin sections by checking nuclear localisation, tissue controls and the catalog antibody’s documented retrieval conditions.
What retrieval conditions should I try when EHMT2 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01055-2). The documented tissue example used 2 μg/ml catalog antibody overnight at 4°C, so compare retrieval changes while holding antibody concentration and incubation constant (datasheet A01055-2; standard IHC practice). Check whether nuclei stain in a previously responsive section alongside the test section, and inspect tissue morphology after heating (standard IHC practice). If staining remains weak, compare a shorter or longer heating interval on matched sections; excessive heating can damage morphology and complicate nuclear scoring (standard IHC practice).
Could fixation explain inconsistent EHMT2 staining between paraffin blocks?
Target-specific sensitivity of EHMT2 to fixation is unknown from the supplied evidence; the catalog tissue caption does not state a fixative (datasheet A01055-2). Record each block’s fixative, fixation duration and processing history, then compare sections processed together with the same pH 8.0 EDTA retrieval (datasheet A01055-2; standard IHC practice). Uneven fixation or processing can alter antigen accessibility and morphology in paraffin sections, so assess both before changing antibody concentration (standard IHC practice). Do not infer a preferred fixative from EHMT2 tissue staining patterns or its nuclear localisation; those observations do not establish fixation sensitivity (HPA tissue IHC; UniProt Q96KQ7; standard IHC practice).
How should I assess cytoplasmic staining when EHMT2 is expected in nuclei?
Prioritise nuclear staining when assessing EHMT2: UniProt places it in the nucleus and euchromatic chromosome regions, while HPA reports nuclear tissue staining (UniProt Q96KQ7; HPA tissue IHC). HPA subcellular images support nucleoplasm and nuclear speckles, so compare the chromogen pattern with a nuclear counterstain at the same cell boundaries (HPA subcellular; standard IHC practice). Score diffuse cytoplasmic colour separately until it is supported by an independent control, particularly if nuclei in the same cells are unstained (standard IHC practice). Review section edges, folds and damaged areas for staining that follows tissue damage rather than intact nuclei (standard IHC practice).
Can isoforms or epitope masking explain different EHMT2 staining patterns?
EHMT2 has 3 listed isoforms and multiple modified residues, including phosphorylation near its amino terminus (UniProt Q96KQ7). The supplied caption does not identify the antibody’s epitope, so it cannot establish which isoforms or modification states this reagent detects (datasheet A01055-2; UniProt Q96KQ7). Compare matched sections under the documented pH 8.0 EDTA retrieval before attributing a staining difference to an isoform (datasheet A01055-2; standard IHC practice). If epitope coverage is essential, obtain the immunogen or epitope specification and use an independently validated reagent or orthogonal assay to test the interpretation (standard IHC practice).
How can IF help check an ambiguous chromogenic EHMT2 pattern?
Use IF/ICC as a separate assay: HPA reports EHMT2 in the nucleoplasm and nuclear speckles, but the supplied IHC caption does not establish IF conditions (HPA subcellular; datasheet A01055-2). For a vaginal epithelial sample, multiplex EHMT2 with a squamous epithelial marker and a nuclear stain to identify the expected cell population (HPA tissue IHC: high in vaginal squamous epithelial cells; standard IF practice). Choose fluorophores after checking tissue autofluorescence, favouring a channel with lower background for the weaker signal (standard IF practice). Because EHMT2 is nuclear and has no transmembrane segment, optimise permeabilisation for nuclear antibody access while preserving morphology (UniProt Q96KQ7; standard IF practice).
How do I reduce diffuse or tissue-specific chromogenic background?
First compare a no-primary control with the complete stain to locate background from detection reagents or endogenous activity (standard IHC practice). The catalog example used biotinylated secondary antibody, streptavidin–biotin detection and DAB, so include a control that can reveal background from that detection system (datasheet A01055-2; standard IHC practice). Apply a peroxidase block before DAB development, then optimise blocking and washing while keeping the documented 2 μg/ml primary condition as a reference (datasheet A01055-2; standard IHC practice). Judge improvement by clearer nuclear staining in intact cells, not simply by reduced overall colour across the section (UniProt Q96KQ7; standard IHC practice).
What is a defensible way to quantify EHMT2 in stained sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis region before scoring, and exclude folds, necrosis and damaged edges (standard IHC practice). EHMT2 is primarily nuclear, so calculate a nuclear H-score from intensity and percentage of positive cells, or report the percentage of positive nuclei (UniProt Q96KQ7; HPA tissue IHC; standard IHC practice). Normalise positive counts to all evaluable nuclei in the same annotated compartment; for spatial counts, report positive nuclei per mm² of evaluable tissue (standard IHC practice). Keep retrieval, detection, counterstain and imaging settings consistent across sections, and report thresholds and excluded regions with the results (standard IHC practice).
Which findings support true EHMT2 staining rather than an artefact?
A credible EHMT2 signal should resolve to intact nuclei and be assessed against the expected cell population; HPA reports high staining in vaginal squamous epithelial cells (UniProt Q96KQ7; HPA tissue IHC). Compare suspicious cytoplasmic colour, edge accentuation or necrotic regions with preserved tissue elsewhere on the section (standard IHC practice). Check a no-primary control for endogenous enzyme or detection-system signal before treating DAB colour as antigen-specific (standard IHC practice). Interpret a negative result cautiously: HPA reports undetected staining in some sampled cell populations, and its tissue IHC profile has supported reliability with medium RNA–staining consistency (HPA tissue IHC).
Boster reagents
Best EHMT2 / Histone-lysine N-methyltransferase EHMT2 IHC Antibodies
A01055-2 has paraffin-section IHC images from human liver cancer, human rectal cancer, and mouse ovary, plus an IF image from A549 cells (catalog image captions).
Real IHC data
Anti-EHMT2/G9A Antibody ®
Cat # A01055-2
A01055-2 is the only card rendered, with its own IHC figure showing a paraffin-embedded human liver cancer section (A01055-2 IHC figure caption). Additional catalog images show IHC in human rectal cancer and mouse ovary sections and IF in A549 cells (A01055-2 image captions).
Which to pick: For tissue IHC, choose A01055-2: its own paraffin-section figure used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A01055-2 IHC figure caption). For IF/ICC, A01055-2 has an A549 IF image at 5 μg/ml, while M01055-1 lists ICC/IF without an image in this payload (A01055-2 IF caption; M01055-1 catalog applications and image list). For cross-species IHC, A01055-2 has human and mouse tissue images and declared rat reactivity; M01055-1 is a rabbit monoclonal with declared human, mouse, and rat reactivity but no IHC image here, while M01055 lists human reactivity only (catalog IHC captions, applications, reactivity, 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 conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Some cell populations have no detectable stain (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unknown (UniProt); Regulation: High 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.