This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
KMT2E / Histone reader KMT2E · IHC design guide
Design Immunohistochemistry for KMT2E
Plan chromogenic KMT2E IHC in paraffin sections using hematopoietic cells in bone marrow as a medium-staining reference (HPA tissue IHC). The catalog antibody lists an IHC dilution range of 1:50–1:200 (datasheet); assess cytoplasmic staining in immune-cell subsets (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KMT2E 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 | Cytoplasm in immune-cell subsets (HPA tissue IHC) | |
| Staining pattern | Subsets of immune cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Bone marrow | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08526) | |
| Caveat | Marrow peroxidase may mimic positive staining (HPA tissue IHC; standard IHC practice) | |
| Regulation | OGT and USP7 stabilize isoform 3 (UniProt) | |
| Isoform / epitope | 8 isoforms; check epitope coverage; no TM sidedness (UniProt) |
Section 1
Recommended KMT2E IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with the published esophageal cancer tumor-section protocol (PMC12527620).
| Sample | Paraffin-embedded rat ovary tissue; fixative not specified (datasheet A08526) |
| Fixation | Image fixative and duration unreported (datasheet A08526); 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-KMT2E, 1:50-1:200 (datasheet A08526) |
| 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 | KMT2E-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in subsets of immune cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval); compare heated citrate retrieval when adapting the published protocol (PMC12527620).
Section 2
What Is the Expected KMT2E Staining Pattern?
KMT2E can localize to the nucleoplasm, cytoplasm and other compartments (UniProt Q8IZD2 subcellular location); it has no transmembrane segment (UniProt Q8IZD2 topology). In tissue IHC, expect cytoplasmic staining in subsets of immune cells, including medium staining in bone marrow hematopoietic cells (HPA tissue IHC). HPA rates the tissue pattern Approved, with external verification pending (HPA tissue IHC reliability).
| Cytoplasmic signal in some bone marrow hematopoietic cells, with unstained neighbors. | This fits the reported medium staining in hematopoietic cells and subset pattern in immune cells (HPA tissue IHC). Score the stained cells and compartment; a whole-section average would obscure the subset pattern (general IHC practice). |
| Predominant nuclear staining, without the expected immune-cell cytoplasmic pattern. | Check the antibody and slide controls before rejecting it: nucleoplasmic localization is supported by ICC-IF (HPA subcellular), while the reported tissue IHC pattern is cytoplasmic (HPA tissue IHC). Uniform nucleolar staining is especially discordant with the reported absence from nucleoli (UniProt Q8IZD2 subcellular location). |
| Strong staining in cells reported as unstained, such as adipocytes or adrenal glandular cells. | These cell types were not detected in the supplied HPA tissue IHC observations (HPA tissue IHC). If staining persists in controls, consider cross-reactivity or endogenous detection activity as general IHC possibilities (general IHC practice); one discrepant slide cannot identify the cause. |
| Color spreads across stroma or slide areas without clear cell boundaries. | Treat diffuse staining as background until controls support a cellular signal (general IHC practice). It does not establish KMT2E expression or a subcellular location; compare tissue morphology, antibody-omission controls and the reported immune-cell subset pattern (HPA tissue IHC; general IHC practice). |
| No detectable signal in bone marrow hematopoietic cells. | The result conflicts with the supplied medium IHC observation (HPA tissue IHC). Check section quality and assay controls, then review the catalog antibody's IHC-P conditions (general IHC practice). Absence of staining alone does not establish absence of protein (general IHC practice). |
💡Expected KMT2E appearanceA supported positive tissue-IHC result is medium cytoplasmic staining in a subset of bone marrow hematopoietic cells (HPA tissue IHC); diffuse background or strong staining confined to reported unstained cell types warrants control-based review (HPA tissue IHC; general IHC practice).
| Tissue-pattern confidence | HPA calls the tissue IHC pattern Approved but says staining involves cells or structures not annotated and awaits external verification (HPA tissue IHC reliability); interpret a new tissue result with its controls (general IHC practice). |
| Cell-specific expression | HPA reports cytoplasmic staining in immune-cell subsets and medium staining in bone marrow hematopoietic cells (HPA tissue IHC). Its low tissue RNA specificity does not predict uniform staining of every cell in a section (HPA tissue IHC RNA specificity; general IHC practice). |
| Compartment and cell cycle | UniProt places KMT2E on interphase chromosomes and at mitotic centrosomes, as well as in nuclear and cytoplasmic compartments (UniProt Q8IZD2 subcellular location). A small atypical cell should be judged against morphology and the surrounding pattern (general IHC practice). |
| Isoforms and membrane interpretation | UniProt lists 8 isoforms and reports tumor-associated expression of NKp44L, while the canonical record has no transmembrane segment (UniProt Q8IZD2 isoforms, tissue specificity and topology). Do not assign a membrane-only stain to a particular isoform without isoform-specific evidence (general IHC practice). |
| Antibody evidence | The supplied antibody records give IHC Approved for HPA056125 and ICC Supported for HPA042866 (HPA antibodies). Those ratings apply to different antibodies and applications; they do not show that the catalog antibody reproduces either pattern (HPA antibodies). |
| IF/ICC Q: Should its pattern match tissue IHC? | A: ICC-IF mainly shows nucleoplasm, with nuclear bodies and cytosol also reported (HPA subcellular); tissue IHC reports cytoplasm in immune-cell subsets (HPA tissue IHC). Compare each result with its own application evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow positive control is blank. | The slide conflicts with reported medium hematopoietic-cell staining (HPA tissue IHC); the failed step is unknown. | Confirm that hematopoietic cells are present, inspect the detection control, and compare retrieval and antibody use with the catalog antibody's IHC-P instructions (general IHC practice). Target-specific retrieval sensitivity is unreported in the supplied evidence. |
| Most cells stain equally, obscuring immune-cell subsets. | Uniform staining conflicts with the reported subset pattern (HPA tissue IHC); background or nonspecific binding is possible (general IHC practice). | Compare an antibody-omission control, inspect cell boundaries, and review blocking, washes and detection conditions (general IHC practice). Score only interpretable cellular staining (general IHC practice). |
| Reported unstained cell types appear positive. | The observation differs from HPA's unstained adipocytes or adrenal glandular cells (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Run appropriate detection controls and compare staining across cell types on the same section (general IHC practice). Treat the discrepancy as unresolved until controlled evidence supports it. |
| Staining appears exclusively nucleolar. | UniProt reports KMT2E absent from the nucleolus (UniProt Q8IZD2 subcellular location). | Verify nuclear boundaries with the counterstain and check background and antibody controls (general IHC practice). Reassess compartment scoring before calling the signal KMT2E. |
| Nuclear ICC-IF and cytoplasmic tissue IHC appear inconsistent. | The supplied applications report different dominant patterns and use different antibody-validation records (HPA subcellular; HPA tissue IHC; HPA antibodies). | Interpret each image within its application and antibody record (HPA antibodies; general IHC practice). Use the separate IF/ICC guide for IF design. |
| A darker section seems to show more KMT2E-positive cells. | Detection intensity or diffuse background can alter apparent scoring (general IHC practice); HPA reports expression in subsets rather than all immune cells (HPA tissue IHC). | Review cell morphology and control staining, then score the fraction and intensity of clearly stained cells consistently across sections (general IHC practice). |
Experiment evidence
IHC & IF images showing KMT2E staining
A08526 · IHCImmunohistochemistry of paraffin-embedded rat ovary using MLL5 antibody at dilution of 1:200 .Perform microwave antigen retrieval with 10 mM
A08526 · IHCImmunohistochemistry of paraffin-embedded human breast cancer using MLL5 antibody at dilution of 1:200 .Perform microwave antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000005483-KMT2E/tissue/cerebral+cortex#imid_16175655
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000005483-KMT2E/tissue/colon#imid_16175589
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000005483-KMT2E/tissue/liver#imid_16175582
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000005483-KMT2E/tissue/kidney#imid_16175591
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA042866; green = KMT2E, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA042866; green = KMT2E, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KMT2E IHC & IF
🧪Run bone marrow first and score hematopoietic cells for staining (HPA: Medium in bone marrow hematopoietic cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the bone marrow slide, treat cells outside the annotated hematopoietic population as internal negatives only if they show background staining, since HPA does not identify a negative cell type there.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KMT2E in RT-4, Rh30, 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 clonality, and a biological negative such as knockout tissue or a peptide-blocked primary antibody if its immunizing peptide is available (standard IHC controls). Quench endogenous peroxidase and inspect unstained bone marrow for pigment or background before interpreting chromogenic signal (standard bone marrow IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A08526 paraffin-section caption does not state a fixative (caption: A08526). That caption reports microwave antigen retrieval in 10 mM PBS, pH 7.2, with primary antibody at 1:200 (caption: A08526); it does not establish that retrieval is required for every specimen. The supplied evidence does not establish whether frozen sections or IF are easier; endogenous peroxidase and pigment can complicate chromogenic scoring in bone marrow (standard bone marrow IHC practice).
HPA tissue IHC evidence for KMT2E
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Antibody staining in cells/structures not annotated, view images. 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KMT2E IHC Tips
Troubleshoot KMT2E staining in paraffin sections by checking retrieval, cell compartment, controls and scoring before interpreting tissue differences.
Which retrieval conditions should I try first for KMT2E IHC?
Start KMT2E paraffin section IHC with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval specification). If staining is weak, test 10 mM PBS, pH 7.2, with microwave retrieval on an adjacent section; that condition accompanied the rat ovary image for catalog antibody A08526 (A08526 tissue-IHC caption). Keep section thickness, antibody dilution and detection conditions matched while comparing retrievals (general IHC practice). Score signal and tissue damage together, because harsher retrieval can increase background or disrupt morphology (general IHC practice). The caption does not report fixation, so its retrieval result cannot establish performance for every preparation (A08526 tissue-IHC caption).
How should I investigate weak staining when fixation history varies?
Record the fixative, fixation duration and processing history for every block before comparing KMT2E staining (general IHC practice). Target-specific fixation sensitivity is unknown here: the selected paraffin section caption does not state a fixative, and the supplied evidence does not test fixation conditions (A08526 tissue-IHC caption). Compare differently processed blocks using adjacent sections, matched retrieval and the same antibody dilution, then judge nuclear signal alongside preserved morphology (general IHC practice; HPA subcellular: supported nucleoplasm). Include a no-primary control to identify detection background before changing retrieval or antibody concentration (general IHC practice). Do not assign a staining difference to fixation alone when specimen handling also differs (general IHC practice).
Should a valid KMT2E stain be nuclear, cytoplasmic or both?
Assess nuclear staining first, since KMT2E is mainly nucleoplasmic and can also occupy nuclear bodies (HPA subcellular: supported nucleoplasm; approved nuclear bodies). Cytoplasmic signal is plausible, but evaluate it against the cell type and a no-primary control: cytosol is an additional supported location, and tissue IHC reports cytoplasmic expression in subsets of immune cells (HPA subcellular: supported cytosol; HPA tissue IHC profile). Exclude nucleoli from the expected pattern because KMT2E is reported absent there (UniProt Q8IZD2: PubMed:14718661). Annotate mitotic cells separately; KMT2E can localize to centrosomes during mitosis (UniProt Q8IZD2: PubMed:23798402). Compare compartments on matched sections before treating diffuse cytoplasmic color as specific (general IHC practice).
Could an epitope or isoform explain discordant KMT2E staining?
Check the catalog antibody's immunogen or mapped epitope against the 8 listed KMT2E isoforms before comparing results across antibodies (UniProt Q8IZD2: isoforms 1–7 and NKp44L). An epitope shared by several isoforms cannot distinguish them by IHC, while an isoform-specific epitope requires an independently documented sequence match (general immunostaining practice). KMT2E contains a SET domain at residues 330–447 and reported phosphorylation sites, but their effects on this antibody's binding are not established here (UniProt Q8IZD2: domains and modified residues). Compare adjacent sections with the same retrieval and detection settings, then verify any compartment difference with an independent, mapped reagent (general IHC practice). Do not infer an isoform solely from stain location (general immunostaining practice).
How can IF help assess a disputed KMT2E IHC pattern?
Use IF on a separate preparation to compare KMT2E with a marker for the cell population identified in the IHC section; bone marrow hematopoietic cells provide a reported tissue-IHC positive population (HPA tissue IHC: medium in hematopoietic cells). Choose spectrally separated fluorophores and inspect unstained tissue in each channel before selecting an emission range, because tissue autofluorescence can mimic signal (general IF practice). KMT2E has no transmembrane segment, so permeabilise fixed cells when testing intracellular nuclear and cytosolic staining, then include a no-primary control (UniProt Q8IZD2: topology; HPA subcellular: nucleoplasm and cytosol; general IF practice). HPA reports IF images in RT-4, Rh30, U2OS and KOLF2.1J cells; their patterns require comparison with the corresponding tissue IHC (HPA subcellular).
What should I change when brown staining appears throughout the section?
First inspect a no-primary section to separate detection-system color from antibody-dependent staining (general IHC practice). Apply an endogenous peroxidase block before chromogenic detection, and compare DAB development times on matched sections if diffuse brown signal persists (general chromogenic IHC practice). Titrate the catalog antibody around its documented 1:200 image condition while keeping retrieval and detection fixed; that dilution was used for paraffin-embedded rat ovary, not established as universal (A08526 tissue-IHC caption). Check whether color follows tissue edges, folds or damaged areas rather than intact cells (general IHC practice). Report nuclear and cytoplasmic background separately because both compartments can contain KMT2E signal (HPA subcellular: nucleoplasm and cytosol).
How should I score KMT2E staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; KMT2E has supported nucleoplasmic and cytosolic locations, and tissue IHC reports cytoplasmic staining in immune-cell subsets (HPA subcellular; HPA tissue IHC profile). For each compartment, record percent positive cells and intensity, or calculate an H-score from the percentages in intensity categories 0–3 (general IHC scoring practice). If the endpoint is immune-cell abundance, report positive-cell density per mm² of viable tissue and normalise to total eligible cells or assessed viable area (general IHC scoring practice). Keep retrieval, development time, imaging and positivity thresholds consistent across samples (general IHC practice). Report nuclear and cytoplasmic results separately so a shift in cell composition does not masquerade as altered intracellular localisation (general IHC scoring practice).
How do I distinguish convincing KMT2E staining from artefact?
Look for staining in intact cells with a defensible compartment: nucleoplasm is the principal supported location, while nuclear bodies and cytosol are additional reported locations (HPA subcellular). A nucleolus-only pattern conflicts with the reported absence of KMT2E from nucleoli, so investigate it before calling it positive (UniProt Q8IZD2: PubMed:14718661). Check cell identity as well as intensity: tissue IHC reports cytoplasmic staining in subsets of immune cells and medium staining in bone marrow hematopoietic cells (HPA tissue IHC profile). Exclude edge effects, necrotic areas and signal present in a no-primary control; check endogenous peroxidase when brown deposits persist (general chromogenic IHC practice). HPA labels its tissue staining Approved pending external verification, so treat an unexpected distribution as provisional until independently confirmed (HPA tissue IHC reliability).
Boster reagents
Best KMT2E / Histone reader KMT2E IHC Antibodies
A08526 has pictured paraffin-section IHC in rat ovary, human breast cancer and mouse lung (catalog IHC captions); IF/ICC is listed, but no IF image is supplied (catalog applications; IF images).
Real IHC data
Anti-MLL5 KMT2E Antibody
Cat # A08526
A08526 will render with its rat ovary paraffin-section IHC figure at 1:200 (catalog IHC caption). Its other captions show paraffin-section IHC in human breast cancer and mouse lung; IF/ICC is listed for Human, Mouse and Rat without an IF figure (catalog IHC captions; applications; reactivity; IF images).
Which to pick: For tissue IHC, choose A08526: its captions document paraffin-section staining after microwave retrieval in 10 mM PBS, pH 7.2, at 1:200 (catalog IHC captions). For IF/ICC, A08526 is the listed option with a 1:50–1:200 range, although no IF image is supplied (catalog applications; IF dilution; IF images). For cross-species work, A08526 lists Human, Mouse and Rat reactivity (catalog reactivity); its IHC captions do not report a fixative, and its clone field is blank (catalog IHC captions; clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IZD2 (KMT2E_HUMAN, Histone reader KMT2E).
- Human Protein Atlas. KMT2E tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KMT2E subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies and cytosol..
- Human Protein Atlas. KMT2E antibody validation summary (2 antibodies).
- ALYREF-Mediated Regulation of TBL1XR1 and KMT2E Synergistically Upregulates APOC1, Contributing to Oxaliplatin Resistance in Esophageal Cancer. Cancer research and treatment 2025 — PMC12527620.
- PubMed PMID:12101424 — UniProt-cited evidence.
- PubMed PMID:23958951 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KMT2E (Histone reader KMT2E). Expected localisation Cytoplasm in immune-cell subsets (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A08526 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat ovary tissue; fixative not specified (datasheet A08526); 2. Fixation: Image fixative and duration unreported (datasheet A08526); 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-KMT2E, 1:50-1:200 (datasheet A08526); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08526); Caveat: Marrow peroxidase may mimic positive staining (HPA tissue IHC; standard IHC practice); Isoform / epitope: 8 isoforms; check epitope coverage; no TM sidedness (UniProt); Regulation: OGT and USP7 stabilize isoform 3 (UniProt). Evidence: UniProt Q8IZD2, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08526, PMC12527620. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KMT2E/immunohistochemistry-ihc-guide.