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KDM8 / Bifunctional peptidase and arginyl-hydroxylase JMJD5 · IHC design guide
Design Immunohistochemistry for KDM8
Plan chromogenic KDM8 IHC on paraffin sections with the catalog antibody at a starting concentration of 2.5 μg/mL (datasheet). Compare observed cytoplasmic staining with tissue IHC evidence while treating nuclear and chromosomal location as a molecular expectation (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KDM8 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 | Cytoplasmic tissue staining (HPA tissue IHC); nuclear/chromosomal molecular location (UniProt) | |
| Staining pattern | General cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue |
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 low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Highly expressed in breast cancer cells (UniProt) | |
| Isoform / epitope | 3 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended KDM8 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with reported KDM8 staining of prostate tissue microarrays and mouse pancreatic tumors (PMC6755995; PMC12622147; PMC12407890).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08840); 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-KDM8, 2.5 μg/mL (datasheet A08840) |
| 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 | KDM8-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. 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 rule; UniProt Q8N371: nucleus).
Section 2
What Is the Expected KDM8 Staining Pattern?
KDM8 is annotated in the nucleus and on chromosomes (UniProt Q8N371), while tissue IHC shows general cytoplasmic staining (HPA tissue IHC) and ICC-IF places it mainly in the nucleoplasm, with additional cytosolic signal (HPA ICC-IF). High IHC staining is reported in several glandular, epithelial, neuronal and muscle cell populations (HPA tissue IHC). Interpret the compartment carefully: HPA rates its tissue IHC Approved but reports low consistency with RNA expression (HPA tissue IHC).
| Cytoplasmic staining in adrenal, salivary or stomach glandular cells, or bronchial respiratory epithelial cells. | This matches cell populations reported as High in tissue IHC (HPA tissue IHC). Record the stained cell type and intensity alongside the compartment: the tissue profile is generally cytoplasmic (HPA tissue IHC), although UniProt places KDM8 in the nucleus and on chromosomes (UniProt Q8N371). |
| Strong staining confined to a compartment that disagrees with both the reported tissue IHC and nucleoplasmic ICC-IF patterns. | Treat an isolated, unexpected compartment as a possible artefact until controls support it (general IHC practice). Nuclear signal alone is not automatically wrong: nucleoplasm is the main ICC-IF location (HPA ICC-IF), and nucleus and chromosome are UniProt locations (UniProt Q8N371). |
| Prominent staining in a cell population reported as undetected, such as adipocytes. | Adipocytes are reported as Not detected (HPA tissue IHC). Check whether the signal belongs to adipocytes or adjacent cells; if it persists in the wrong cell type, consider cross-reactivity or endogenous detection activity (general IHC practice). An unexpected result is not, by itself, proof of either cause. |
| Diffuse color across cells, extracellular areas and the slide, with little distinction between stained and unstained structures. | This is difficult to score as cell-specific KDM8 staining (general IHC practice). Check the no-primary control and assess blocking, washes and chromogen development (general IHC practice); the HPA tissue pattern does not establish a target-specific cause for diffuse background (HPA tissue IHC). |
| No visible staining in a section expected to contain a reported High cell population. | First check that the relevant cells are present: cardiomyocytes, cerebral cortical neuronal cells and splenic white-pulp cells are among the High populations (HPA tissue IHC). An absent signal calls for a run-control check before concluding biological absence (general IHC practice), especially given the reported low IHC–RNA consistency (HPA tissue IHC). |
💡Expected KDM8 appearanceA plausible positive IHC result is discernible, predominantly cytoplasmic color in a reported High cell population, such as bronchial respiratory epithelium (HPA tissue IHC); diffuse color or convincing adipocyte staining warrants control review (general IHC practice; HPA tissue IHC: adipocytes Not detected).
| Compartment differs by assay | Tissue IHC is described as generally cytoplasmic (HPA tissue IHC); ICC-IF is mainly nucleoplasmic with additional cytosol (HPA ICC-IF), and UniProt lists nucleus and chromosome (UniProt Q8N371). Report the method when describing location. |
| Cell type matters more than a whole-tissue label | High staining is assigned to specified cells within tissues, including cerebellar granular-layer cells and splenic white-pulp cells (HPA tissue IHC). Sample composition therefore matters when selecting a comparison field (general IHC practice). |
| Strength of tissue evidence | The tissue IHC reliability label is Approved, with low consistency between antibody staining and RNA expression (HPA tissue IHC). The listed High and Not detected calls describe observed staining; they should be weighed with slide controls rather than treated as independent proof of specificity. |
| Antibody validation scope | HPA026545 is Approved for IHC and Supported for ICC (HPA antibodies). The supplied record does not assign it Enhanced IHC validation (HPA antibodies); do not infer independent reproduction of this tissue pattern from the listed status. |
| Isoforms and epitope uncertainty | UniProt lists three KDM8 isoforms and a JmjC domain at residues 271–416 (UniProt Q8N371). The supplied evidence gives no antibody epitope or isoform coverage, so a difference in staining cannot be assigned to a particular isoform. |
| Protein topology and processing | KDM8 has no transmembrane segment, signal peptide, propeptide or listed glycosylation site, and its annotated chain spans residues 1–416 (UniProt Q8N371). These annotations do not establish how fixation or antigen retrieval affects IHC signal. |
| Situation | Likely cause | Next action |
|---|---|---|
| No color appears in a reported High cell population. | The expected cells may be missing from the section, or the staining run may have failed (general IHC practice); HPA reports High staining only for specified cell types (HPA tissue IHC). | Confirm the cell population morphologically, then review the run control, primary-antibody step, detection reagents and development (general IHC practice). Do not use tissue RNA enrichment alone to decide whether the stain succeeded: IHC–RNA consistency is low (HPA tissue IHC). |
| Color appears broadly, including outside recognizable cells. | Non-specific reagent binding, incomplete washes or overdevelopment can produce diffuse chromogenic background (general IHC practice). | Compare with a no-primary control; review blocking, wash steps and development time (general IHC practice). Score KDM8 only where cell boundaries and a reproducible compartment can be assessed (general IHC practice). |
| A high-intensity signal appears in adipocytes. | Adipocytes are reported as Not detected (HPA tissue IHC); an unexpected signal could reflect misidentified adjacent cells, cross-reactivity or endogenous detection activity (general IHC practice). | Verify cell identity on the counterstained section and compare a no-primary control (general IHC practice). If an enzyme-based chromogen is used, check the corresponding endogenous-activity control (general IHC practice). |
| Only nuclei stain in a tissue IHC section. | Nuclear staining agrees with UniProt location and the main ICC-IF location (UniProt Q8N371; HPA ICC-IF), but differs from HPA's general cytoplasmic tissue IHC profile (HPA tissue IHC). | Document the compartment and cell type, compare control sections and avoid calling the result either confirmed or artifactual from location alone (general IHC practice). |
| Cytoplasmic color is present, but the expected High cells are hard to distinguish. | The High calls refer to particular populations, such as glandular cells in adrenal gland, salivary gland and stomach (HPA tissue IHC); surrounding cells can complicate visual attribution (general IHC practice). | Use morphology and the counterstain to identify the reported population, then score its staining separately from neighboring cells and background (general IHC practice). |
| IF/ICC Q: Is cytosolic KDM8 signal compatible with the reference pattern? | A: Yes. HPA reports an additional cytosolic location alongside its approved main nucleoplasmic location (HPA ICC-IF); UniProt lists nucleus and chromosome (UniProt Q8N371). | Describe nucleoplasmic and cytosolic signal separately and compare appropriate imaging controls (general IF practice). Do not transfer the tissue IHC intensity calls directly to IF/ICC images, because those calls come from tissue IHC (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing KDM8 staining
A08840 · IHCImmunohistochemistry of JMJD5 in rat liver tissue with JMJD5 antibody at 2.5 μg/mL.
A08840 · IFImmunofluorescence of JMJD5 in Rat Liver cells with JMJD5 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000155666-KDM8/tissue/cerebral+cortex#imid_6518392
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000155666-KDM8/tissue/stomach+1#imid_6517788
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000155666-KDM8/tissue/colon#imid_6517814
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000155666-KDM8/tissue/liver#imid_6517799
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026545; green = KDM8, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026545; green = KDM8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KDM8 IHC & IF
🧪Run adrenal gland first: its glandular cells must stain (HPA: High in adrenal gland glandular cells). Run adipose tissue as the negative: adipocytes should be undetected (HPA: Not detected in adipocytes); on the adrenal slide, assess any neighboring cells without specific staining as internal background controls without assuming a particular cell type is KDM8-negative (standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KDM8 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and irrelevant IgG controls matched to the primary antibody’s host species and immunoglobulin class, with a matching monoclonal or polyclonal format where feasible; use KDM8-knockout material as a biological specificity control if available (standard IHC practice). On adrenal sections, check endogenous peroxidase background, and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A08840 rat-liver IHC caption does not state a fixative (caption: fixative not stated). Retrieval dependence and whether frozen sections or IF would be easier than paraffin IHC are unreported in the supplied evidence; optimize retrieval empirically on paired sections (standard IHC practice). Interpret adrenal staining against the peroxidase and, where applicable, biotin background controls (standard IHC practice).
HPA tissue IHC evidence for KDM8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KDM8 IHC Tips
Use compartment-aware controls to troubleshoot KDM8 staining in paraffin-section chromogenic IHC; assess IF/ICC separately.
What retrieval should I try when KDM8 staining is weak in paraffin sections?
Use Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 min (page IHC protocol). Keep section thickness, cooling time, antibody concentration and detection conditions constant while comparing retrieved sections with an omitted-primary control (standard IHC practice). If staining remains weak, test an alternative retrieval condition on adjacent sections and judge both signal and tissue preservation before adopting it (standard IHC practice). Score nuclear and cytoplasmic staining separately because UniProt places KDM8 in the nucleus and on chromosomes, while HPA reports approved nucleoplasmic and additional cytosolic localisation (UniProt Q8N371; HPA subcellular).
How can I troubleshoot KDM8 staining that varies between tissue blocks?
The supplied KDM8 tissue-IHC caption does not state a fixative, so target-specific fixation sensitivity is unknown (A08840 caption). Record each block’s documented fixation and processing history, then compare adjacent sections using the same retrieval, antibody incubation and chromogenic detection conditions (standard IHC practice). Include a block that has produced interpretable staining in the same run, and check morphology alongside signal before attributing a difference to KDM8 abundance (standard IHC practice). Avoid using HPA tissue patterns or the UniProt topology record to infer a preferred fixative; neither supplies a KDM8 fixation comparison (HPA tissue IHC; UniProt Q8N371).
How should I assess predominantly cytoplasmic KDM8 staining?
Score nuclear and cytoplasmic staining independently: UniProt places KDM8 in the nucleus and on chromosomes, whereas HPA reports approved nucleoplasmic and additional cytosolic localisation (UniProt Q8N371; HPA subcellular). General cytoplasmic tissue expression is reported by HPA, but its tissue-IHC annotation has low consistency with RNA expression data (HPA tissue IHC). Compare staining within intact cells and against an omitted-primary control, paying particular attention to diffuse signal over tissue edges or damaged areas (standard IHC practice). If only cytoplasmic signal persists, document its distribution and seek independent antibody or orthogonal evidence before assigning it to KDM8 (standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent KDM8 staining?
UniProt lists three KDM8 isoforms and places the JmjC domain at residues 271–416, but the supplied evidence does not identify this antibody’s epitope (UniProt Q8N371; supplied antibody evidence). Check the antibody’s documented immunogen or epitope before assuming that staining represents every isoform (standard IHC practice). Compare adjacent sections under the page retrieval condition while keeping detection settings fixed, since tissue processing and retrieval can alter epitope accessibility (page IHC protocol; standard IHC practice). Report the antibody identity and retrieval method with any compartment-specific result so others can assess which isoform or epitope interpretation remains uncertain (standard IHC practice).
How should I check KDM8 localisation by multiplex IF?
For a separate IF/ICC experiment, pair KDM8 with an independently validated marker of the cell type being assessed and include a nuclear counterstain (standard IF practice). HPA reports approved nucleoplasmic localisation with additional cytosolic localisation and provides ICC/IF images in A-431, U-251MG and U2OS cells (HPA subcellular). Choose fluorophores after inspecting unstained tissue autofluorescence, favouring a spectrally separated far-red channel when shorter-wavelength background is strong (standard IF practice). Because KDM8 is intracellular and has no transmembrane segment, assess fixation-compatible permeabilisation for access to nuclear and cytosolic epitopes; no KDM8-specific IF fixation condition is supplied (UniProt Q8N371; supplied IF evidence; standard IF practice).
What should I check when chromogenic KDM8 staining is widespread?
Run omitted-primary and detection-only controls to identify background from secondary reagents or the chromogenic workflow (standard IHC practice). For peroxidase-based detection, check that the peroxidase block is effective and compare the timing of DAB development across sections (standard IHC practice). Titrate the antibody around a documented starting condition only within its stated application: the A08840 image caption reports 2.5 µg/mL in rat liver tissue IHC without stating fixation (A08840 caption). Interpret widespread cytoplasmic colour cautiously because HPA reports general cytoplasmic tissue expression while also noting low consistency between antibody staining and RNA data (HPA tissue IHC).
How should I quantify KDM8 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, exclude damaged or poorly preserved areas, and apply the same image acquisition settings throughout a comparison (standard IHC practice). Report the percentage of positive cells and staining intensity separately for nuclei and cytoplasm; an H-score combines percentage and intensity on a 0–300 scale (standard IHC practice). For spatial comparisons, report positive-cell density per mm² and normalise it to the analysed tissue area or relevant cell count (standard IHC practice). State the threshold, compartment and sampled regions because UniProt and HPA support different aspects of KDM8 localisation (UniProt Q8N371; HPA subcellular).
When is a KDM8-positive IHC result convincing?
Look for reproducible staining in intact cells with a plausible compartment pattern: UniProt reports nuclear and chromosomal KDM8, and HPA supports nucleoplasmic with additional cytosolic localisation (UniProt Q8N371; HPA subcellular). Check cell identity against the section’s morphology; HPA reports high staining in several specified cell populations, but flags low consistency between its antibody staining and RNA data (HPA tissue IHC). Treat staining confined to section edges, necrotic regions or endogenous peroxidase-rich areas as suspect until controls resolve it (standard IHC practice). A convincing assignment combines appropriate controls, preserved morphology and independent evidence rather than colour intensity alone (standard IHC practice).
Boster reagents
Best KDM8 / Bifunctional peptidase and arginyl-hydroxylase JMJD5 IHC Antibodies
A08840 has real IHC data from rat liver tissue and IF data from rat liver cells (IHC and IF image captions); human, mouse and rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-JMJD5 KDM8 Antibody
Cat # A08840
A08840 is listed for IHC-P and shown staining rat liver tissue at 2.5 μg/mL (catalog applications; IHC image caption). A08840 is also listed for IF and shown in rat liver cells at 20 μg/mL (catalog applications; IF image caption).
Which to pick: Choose A08840 for paraffin-section tissue IHC based on its IHC-P listing and rat liver tissue image; the fixative is unreported (catalog applications; IHC image caption). For IF/ICC, A08840 has an IF image in rat liver cells, while ICC-specific validation is unreported (catalog applications; IF image caption). A08840 is listed as reactive with human, mouse and rat, but the supplied IHC and IF images show rat samples only; its host is rabbit and clonality is unreported (catalog reactivity and host; IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N371 (KDM8_HUMAN, Bifunctional peptidase and arginyl-hydroxylase JMJD5).
- Human Protein Atlas. KDM8 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KDM8 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. KDM8 antibody validation summary (1 antibodies).
- KDM8/JMJD5 as a dual coactivator of AR and PKM2 integrates AR/EZH2 network and tumor metabolism in CRPC. Oncogene 2019 — PMC6755995.
- Allyl Isothiocyanate Suppresses the Proliferation in Oral Squamous Cell Carcinoma via Mediating the KDM8/CCNA1 Axis. Biomedicines 2023 — PMC10604360.
- Hypoxia-induced metastatic heterogeneity in pancreatic cancer. Research square 2025 — PMC12622147.
- Hypoxia-induced metastatic heterogeneity in pancreatic cancer. bioRxiv : the preprint server for biology 2025 — PMC12407890.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KDM8 (Bifunctional peptidase and arginyl-hydroxylase JMJD5). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear/chromosomal molecular location (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A08840 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 A08840); 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-KDM8, 2.5 μg/mL (datasheet A08840); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; check antibody epitope coverage (UniProt); Regulation: Highly expressed in breast cancer cells (UniProt). Evidence: UniProt Q8N371, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08840, PMC6755995, PMC12622147. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KDM8/immunohistochemistry-ihc-guide.