This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
EZH1 / Histone-lysine N-methyltransferase EZH1 · IHC design guide
Design Immunohistochemistry for EZH1
Plan chromogenic EZH1 IHC in paraffin sections with nuclear staining as the expected pattern (HPA tissue IHC). Start the IHC-validated antibody A02494 at 5 μg/mL (datasheet), using colon endothelial cells or duodenal Brunner glands as positive tissue controls (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EZH1 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 | Nucleus (UniProt) | |
| Staining pattern | Nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Colon+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 and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 5 isoforms; epitope coverage unknown (UniProt) |
Section 1
Recommended EZH1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is presented alongside four published EZH1 IHC protocols (PMC8698684; PMC9285492; PMC7640435; PMC6743615).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02494); 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-EZH1, 5 μg/mL (datasheet A02494) |
| 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 | EZH1-positive staining in endothelial cells of colon (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min for nuclear EZH1 (page retrieval rule; UniProt Q92800). Citrate retrieval is a published alternative (PMC9285492).
Section 2
What Is the Expected EZH1 Staining Pattern?
EZH1 should appear mainly in nuclei across many tissues (HPA tissue IHC: nuclear expression in most tissues; UniProt Q92800: nucleus). The clearest supplied IHC examples are colon endothelial cells and duodenal Brunner glands, both scored High (HPA tissue IHC). HPA rates the tissue profile Supported, with medium consistency between staining and RNA data (HPA tissue IHC). EZH1 has no transmembrane segment (UniProt Q92800 topology).
| Distinct nuclear staining in colon endothelial cells or duodenal Brunner glands. | This matches the supplied High IHC observations in those cell populations (HPA tissue IHC). Assess the named cells rather than assigning one score to the whole tissue; neighboring cells may differ. A nuclear pattern also agrees with the annotated EZH1 location (UniProt Q92800: nucleus). |
| Predominantly cytoplasmic, membranous, or extracellular color, with little nuclear signal. | This conflicts with nuclear tissue staining (HPA tissue IHC) and nuclear localization (UniProt Q92800). Treat the compartment mismatch as a possible staining artefact; review the control slide, detection background, and morphology before calling it EZH1. The absence of a transmembrane segment gives no basis for an expected membrane pattern (UniProt Q92800 topology). |
| Strong staining in a population listed as Not detected, such as adipocytes or cardiomyocytes. | That result conflicts with those specific HPA observations (HPA tissue IHC: adipocytes and cardiomyocytes Not detected). Consider cross-reactivity or endogenous detection activity, especially if the expected nuclear pattern is absent. A Not detected score applies to the named cells in the sampled tissue; it does not establish that every cell there lacks EZH1. |
| Color spreads across tissue or pools outside identifiable nuclei. | Diffuse color is difficult to score as EZH1 when the reference pattern is nuclear (HPA tissue IHC; UniProt Q92800). Check whether a no-primary control also develops color, then inspect blocking, washes, and chromogen development (general IHC practice). Interpret only signal that can be assigned to intact cells and their nuclei. |
| No convincing nuclear signal in colon endothelial cells or duodenal Brunner glands. | These are the supplied High examples (HPA tissue IHC), so a blank result warrants a run-level check before a biological conclusion. Confirm that the chosen antibody is validated for IHC and that a positive control worked; HPA lists HPA077684 as IHC Supported (HPA antibody validation). Repeat with adjusted general IHC conditions if controls fail. |
💡Expected EZH1 appearanceCall a positive result when identifiable nuclei in colon endothelial cells or duodenal Brunner glands show clear, relatively strong chromogenic staining (HPA tissue IHC: both High; UniProt Q92800: nucleus); diffuse extracellular color or dominant membrane staining is suspect against that reference pattern (HPA tissue IHC; UniProt Q92800 topology).
| Cell population and tissue context | HPA scores colon endothelial cells and duodenal Brunner glands High, several other named populations Medium or Low, and selected populations Not detected (HPA tissue IHC). Choose and score the specified cell population; an entire organ is an imprecise positive or negative control. |
| Antibody evidence | The tissue profile is Supported, with medium consistency between antibody staining and RNA expression, and HPA notes staining in cells or structures that are not annotated (HPA tissue IHC). HPA077684 is IHC Supported; HPA005478 is ICC Supported, with no IHC status supplied for it (HPA antibody validation). |
| Cellular compartment and complex | EZH1 is nuclear and colocalizes with H3K27me3 (UniProt Q92800); ICC-IF places it in the nucleoplasm with Supported status (HPA subcellular). These observations guide compartment scoring. H3K27me3 localization alone does not identify an EZH1-positive cell. |
| Isoforms and antigen recognition | UniProt lists 5 EZH1 isoforms (UniProt Q92800). The payload supplies no epitope position or isoform coverage for the IHC antibody, so do not infer which variants contribute to staining or explain a negative slide by assuming a particular variant. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control nuclei are blank. | The run may have insufficient detectable signal; colon endothelial cells and duodenal Brunner glands are High reference populations (HPA tissue IHC). | Check the antibody's IHC validation, control-slide handling, retrieval and detection settings against the applicable IHC procedure (general IHC practice). Avoid attributing the blank slide to EZH1-specific fixation sensitivity; none is supplied. |
| The sample is weak but a control is clearly nuclear. | Signal may be lower in the sampled cell population; HPA records Medium and Low populations as well as High ones (HPA tissue IHC). | Identify the actual cell population and compare like with like. Score nuclear signal against the working positive control and morphology; do not use tissue-wide absence as a cell-level finding (general IHC practice). |
| Cytoplasmic or membrane color dominates. | The distribution disagrees with the nuclear EZH1 reference (HPA tissue IHC; UniProt Q92800: nucleus). | Check a no-primary control and the detection reagents, then review whether nuclear counterstain and tissue morphology permit compartment scoring (general IHC practice). Do not score the misplaced color as a positive EZH1 result. |
| Background obscures nuclei throughout the section. | Nonspecific reagent binding or endogenous detection activity can produce broad chromogenic background (general IHC practice). | Compare no-primary and detection controls; review blocking, washing, and chromogen development (general IHC practice). Reassess only identifiable nuclear staining, consistent with the HPA tissue profile (HPA tissue IHC). |
| A listed Not detected population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA lists adipocytes and cardiomyocytes as Not detected in their respective tissues (HPA tissue IHC). | Check localization and control-slide background before interpreting the discrepancy. Record the exact stained cell population and avoid extending that HPA score to every cell in the tissue. |
| Can IF/ICC help assess the compartment? | HPA reports Supported nucleoplasmic localization by ICC-IF and lists A-431, U-251MG, and U2OS images (HPA subcellular). | Use that observation as a localization cross-check only. Interpret the paraffin IHC result against its own nuclear tissue pattern and IHC antibody evidence (HPA tissue IHC; HPA antibody validation). |
Experiment evidence
IHC & IF images showing EZH1 staining
A02494 · IHCImmunohistochemistry of EZH1 in human lung tissue with EZH1 antibody at 5 μg/mL.
A02494 · IFImmunofluorescence of EZH1 in human lung tissue with EZH1 antibody at 20 μg/mL.
A02494-1 · IFIF analysis of EZH1 using anti-EZH1 antibody (A02494-1) and anti-Tubulin Alpha antibody (M03989-3). EZH1 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000108799-EZH1/tissue/cerebral+cortex#imid_20380837
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000108799-EZH1/tissue/colon#imid_20384867
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000108799-EZH1/tissue/liver#imid_20384884
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000108799-EZH1/tissue/kidney#imid_20384877
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005478; green = EZH1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005478; green = EZH1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EZH1 IHC & IF
🧪Run colon first and score endothelial cell nuclei for EZH1 staining (HPA: High in colon endothelial cells; UniProt Q92800: nucleus). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the colon slide, assess surrounding non-endothelial cells for background without assuming they are EZH1-negative (HPA: colon result specifies endothelial cells only).
Positive control tissue: Colon (Endothelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show EZH1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched isotype control chosen to match the primary antibody’s host species and clonality (standard IHC practice). Use EZH1 knockout tissue or a validated immunizing-peptide block as a biological specificity control, and block endogenous peroxidase before chromogenic detection while checking the colon slide for residual background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied EZH1 evidence; optimize antigen retrieval with the catalog antibody’s IHC-P protocol (supplied EZH1 evidence; standard IHC practice). The selected A02494 human lung IHC caption reports 5 µg/mL but leaves the fixative unreported (selected A02494 IHC caption). The supplied evidence does not establish that frozen sections or IF are easier for EZH1; in colon, assess residual peroxidase staining as a possible chromogenic background artefact (supplied EZH1 evidence; standard IHC practice).
HPA tissue IHC evidence for EZH1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glands of Brunner | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Cervix | 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 → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced EZH1 IHC Tips
Troubleshoot EZH1 staining in paraffin sections by checking nuclear localisation, retrieval, controls and cell specific scoring (UniProt Q92800; HPA tissue IHC).
What retrieval conditions should I try when EZH1 nuclear staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval specification). Allow sections to cool in retrieval buffer, then compare a known positive section and a no-primary control within the same staining run (standard IHC practice). If nuclei remain weak, adjust heating time in small increments while watching for tissue damage; keep antibody concentration and detection conditions fixed during that comparison (standard IHC practice). Assess signal within nuclei, since EZH1 is nuclear and its supported subcellular location is the nucleoplasm (UniProt Q92800; HPA subcellular).
How can I assess whether fixation is suppressing EZH1 staining?
Target specific fixation sensitivity for EZH1 is unknown from the supplied evidence (provided EZH1 evidence payload). Record the fixative, fixation interval and section processing for each specimen, then compare sections processed under documented conditions using identical retrieval and detection settings (standard IHC practice). The selected antibody image shows EZH1 staining in human lung tissue at 5 µg/mL, but its caption does not state the fixative (caption A02494). If only one batch stains weakly, inspect its morphology and nuclear counterstain before attributing the difference to EZH1 abundance; use a similarly processed positive control to separate processing variation from biological variation (standard IHC practice).
Should EZH1 staining appear outside the nucleus?
Score crisp nuclear staining as the expected pattern: EZH1 is annotated in the nucleus, and supported imaging places it in the nucleoplasm (UniProt Q92800; HPA subcellular). Its CXC region spans residues 504–606 and SET region spans 613–728; neither annotation implies a membrane staining pattern (UniProt Q92800). If cytoplasm or cell borders dominate, compare the no-primary control, shorten chromogen development and inspect whether the signal persists within intact nuclei (standard IHC practice). Keep the nuclear counterstain light enough to distinguish chromogen from chromatin, and document discordant compartments instead of counting them as EZH1 positive cells (standard IHC practice).
Could an isoform or masked epitope explain inconsistent EZH1 staining?
EZH1 has 5 annotated isoforms, so check which sequence region the antibody recognises before comparing staining across samples (UniProt Q92800). The supplied caption establishes tissue staining for SKU A02494 but does not identify its epitope or demonstrate equal recognition of every isoform (caption A02494). Its annotated CXC and SET regions occupy residues 504–606 and 613–728, respectively; use that map only after obtaining the antibody epitope information (UniProt Q92800; standard IHC practice). When nuclear signal varies, compare matched retrieval conditions and a second antibody with a documented, distinct epitope if available; agreement strengthens an interpretation but does not establish isoform identity (standard IHC practice).
How should I plan EZH1 immunofluorescence alongside a cell type marker?
Treat IF/ICC as a separate validation task: the selected SKU caption documents tissue IHC staining, while the supplied HPA subcellular record supports nucleoplasmic localisation by ICC/IF (caption A02494; HPA subcellular). Pair a nuclear EZH1 channel with a marker for the cell type being evaluated, then confirm that both signals belong to the same cell rather than adjacent cells (standard IF practice). Choose fluorophores after measuring tissue autofluorescence, favouring a spectrally separated, brighter channel for weak EZH1 signal (standard IF practice). Because EZH1 is nuclear and has no transmembrane segment, use controlled permeabilisation to expose intracellular epitopes and compare it with a no-primary control (UniProt Q92800; standard IF practice).
What should I change when EZH1 chromogen covers the whole section?
First inspect the no-primary control and the distribution of precipitate; widespread staining there points to detection or endogenous enzyme background (standard IHC practice). For peroxidase detection, verify the peroxidase block and shorten DAB development before changing the EZH1 antibody concentration (standard IHC practice). Increase blocking or washing if diffuse background persists, and titrate the primary around the documented 5 µg/mL image concentration without treating that caption as a universal optimum (caption A02494; standard IHC practice). Interpret discrete nuclear staining separately from pigment, folds and damaged edges, because the supported EZH1 pattern is nucleoplasmic (HPA subcellular; standard IHC practice).
How should I score EZH1 across differently composed sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, then count only staining in intact nuclei as EZH1 signal (HPA subcellular; standard IHC practice). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score using the same intensity thresholds across slides (standard IHC practice). Normalise counts to the number of eligible nuclei in each cell population, or report positive nuclei per mm² with the analysed area stated (standard IHC practice). Record staining run, retrieval and chromogen settings alongside the score, since the HPA tissue profile includes different cell types and staining levels across tissues (HPA tissue IHC; standard IHC practice).
How can I distinguish genuine EZH1 staining from tissue artefact?
A credible positive result is nuclear staining in intact cells: EZH1 is nuclear, and the supported subcellular location is the nucleoplasm (UniProt Q92800; HPA subcellular). Check the cell type as well as the compartment; HPA reports high staining in colon endothelial cells and undetected staining in adipose tissue adipocytes (HPA tissue IHC). Exclude signal concentrated at section edges, folds or necrotic areas, and compare suspicious deposits with a no-primary control for endogenous enzyme activity (standard IHC practice). Interpret a negative field cautiously because HPA rates its tissue IHC evidence as Supported with medium agreement between antibody staining and RNA expression data (HPA tissue IHC).
Boster reagents
Best EZH1 / Histone-lysine N-methyltransferase EZH1 IHC Antibodies
Anti-EZH1 antibodies have IHC images from human lung tissue (A02494 image caption) and IF images from human lung tissue and HeLa cells (A02494 and A02494-1 image captions).
Real IHC data
Anti-EZH1 Antibody
Cat # A02494
Real IF data
Anti-EZH1 Antibody ®
Cat # A02494-1
A02494 lists IHC-P and IF applications, Human, Mouse and Rat reactivity, and images of human lung tissue for both methods (catalog; A02494 image captions). A02494-1 lists ICC and IF applications, Human, Mouse and Rat reactivity, and an IF image of HeLa cells (catalog; A02494-1 image caption).
Which to pick: Choose A02494 for paraffin-section IHC because IHC-P is listed and human lung tissue IHC is shown at 5 μg/mL (catalog; A02494 IHC image caption); the caption does not report a fixative. Choose A02494-1 for IF/ICC because those applications are listed and HeLa cell IF is shown at 5 μg/mL (catalog; A02494-1 IF image caption). For cross-species work, both list Human, Mouse and Rat reactivity, while the supplied IHC and IF images show human samples (catalog; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92800 (EZH1_HUMAN, Histone-lysine N-methyltransferase EZH1).
- Human Protein Atlas. EZH1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. EZH1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. EZH1 antibody validation summary (2 antibodies).
- Divergent Effects of EZH1 and EZH2 Protein Expression on the Prognosis of Patients with T-Cell Lymphomas. Biomedicines 2021 — PMC8698684.
- Downregulation of the enhancer of zeste homolog 1 transcriptional factor predicts poor prognosis of triple-negative breast cancer patients. PeerJ 2022 — PMC9285492.
- Involvement of Hdac3-mediated inhibition of microRNA cluster 17-92 in bronchopulmonary dysplasia development. Molecular medicine (Cambridge, Mass.) 2020 — PMC7640435.
- Targeting EZH1/2 induces cell cycle arrest and inhibits cell proliferation through reactivation of p57(CDKN1C) and TP53INP1 in mantle cell lymphoma. Cancer biology & medicine 2019 — PMC6743615.
- PubMed PMID:8921387 — UniProt-cited evidence.
- PubMed PMID:9473645 — UniProt-cited evidence.
- PubMed PMID:9205841 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EZH1 (Histone-lysine N-methyltransferase EZH1). Expected localisation Nucleus (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A02494 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Endothelial 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 A02494); 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-EZH1, 5 μg/mL (datasheet A02494); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; epitope coverage unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q92800, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A02494, PMC8698684, PMC9285492. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EZH1/immunohistochemistry-ihc-guide.