KDM4A / Lysine-specific demethylase 4A · IHC design guide

Design Immunohistochemistry for KDM4A

Plan paraffin section KDM4A IHC with nuclear localisation in mind (UniProt) while evaluating the reported cytoplasmic tissue staining (HPA tissue IHC). The catalog antibody A02850-1 has a documented IHC dilution of 2–5 μg/mL (datasheet A02850-1); score nuclear and cytoplasmic signal separately.

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for KDM4A (IHC for KDM4A): expected localisation Cytoplasmic; nuclear in neurons and fallopian glands (HPA tissue IHC), antibody A02850-1, validated IHC image, and IHC protocol steps
Printable KDM4A IHC protocol sheet — expected localisation Cytoplasmic; nuclear in neurons and fallopian glands (HPA tissue IHC), antibody A02850-1, controls and protocol steps. Open the full KDM4A IHC guide →

KDM4A 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; nuclear in neurons and fallopian glands (HPA tissue IHC)
Staining pattern Cytoplasm across tissues; neuronal and fallopian gland nuclei (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet A02850-1)
Positive control ⓘ Caudate+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). Selected-image fixative and duration unreported (datasheet A02850-1); verify before use.
Caveat Reported tissue staining has uncertain reliability (HPA tissue IHC)
Regulation FBXO22 promotes KDM4A ubiquitination (UniProt)
Isoform / epitope 2 isoforms; epitope differences unspecified (UniProt)
Section 1

Recommended KDM4A IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published KDM4A methods for endometrial tissues, mouse xenografts, and paraffin specimens (PMC4741634; PMC8417295).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human ovarian serous carcinoma tissue; fixative not specified (datasheet A02850-1)
FixationImage fixative and duration unreported (datasheet A02850-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A02850-1); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A02850-1)
Primary antibodyRabbit anti-KDM4A, 2-5 μg/ml (datasheet A02850-1)
Primary incubationOvernight at 4 °C (datasheet A02850-1)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet A02850-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultKDM4A-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues. Nuclear expression in neuronal cells and glandular cells of fallopian tube. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A02850-1; PMC4741634), then optimize the primary antibody dilution for your specimen.
Section 2

What Is the Expected KDM4A Staining Pattern?

KDM4A is a nuclear histone demethylase with no transmembrane segment (UniProt O75164). In tissue IHC, HPA reports cytoplasmic staining across tissues, with nuclear staining in neuronal cells and fallopian tube glandular cells (HPA: tissue IHC). High staining is reported in caudate and cerebral cortex neurons and duodenal glandular cells (HPA: tissue IHC). Treat compartment calls cautiously: the tissue profile is rated Uncertain (HPA: reliability).

What am I looking at on my slide?
High staining in caudate or cerebral cortex neurons, or duodenal glandular cells; cytoplasmic staining may accompany neuronal nuclear staining (HPA: tissue IHC).This resembles the reported tissue pattern, but agreement alone does not establish antibody specificity. HPA rates the tissue IHC profile Uncertain because antibody staining and RNA expression have medium consistency and external verification is pending (HPA: reliability).
A predominantly nuclear result, or a wholly cytoplasmic result, seems inconsistent with the other source (UniProt O75164; HPA: tissue IHC).UniProt assigns KDM4A to the nucleus, whereas HPA reports widespread cytoplasmic tissue staining and nuclear staining in selected cells. Review cell boundaries and controls before calling either compartment an artefact; these sources do not resolve the discrepancy (UniProt O75164; HPA: tissue IHC).
Strong staining appears in adipocytes, pancreatic exocrine glandular cells, or smooth muscle cells (HPA: Not detected in these cells).These cells are poor positive comparators in HPA tissue IHC. Unexpected signal raises questions about cross-reactivity, endogenous chromogen activity, or scoring, but the Uncertain HPA profile does not make their staining proof of a false positive (HPA: tissue IHC; standard IHC practice).
Color covers tissue broadly without clear cell boundaries or an interpretable intracellular pattern (standard IHC practice).Diffuse background can obscure both the reported cytoplasmic staining and the more restricted nuclear staining. Examine the negative detection control and background in surrounding tissue before assigning a KDM4A score (HPA: tissue IHC; standard IHC practice).
No signal appears in caudate neurons or duodenal glandular cells, where HPA reports High staining (HPA: tissue IHC).Check whether tissue, detection, and controls worked before interpreting absence as biological. HPA's Uncertain reliability means a negative result in one section cannot by itself settle the expected KDM4A pattern (HPA: reliability; standard IHC practice).
💡Expected KDM4A appearanceA plausible positive IHC result shows High staining in caudate neurons or duodenal glandular cells, with the reported cytoplasmic pattern and possible neuronal nuclear staining; broad uninterpretable color or strong signal in HPA Not detected cells warrants investigation (HPA: tissue IHC; standard IHC practice).
How each factor affects the staining
Tissue and cell selection (HPA: tissue IHC)Caudate neurons and duodenal glandular cells are reported High; adipocytes and pancreatic exocrine glandular cells are Not detected. Compare the named cell populations, since a whole-section score can hide their different staining levels (HPA: tissue IHC; standard IHC practice).
Compartment interpretation (UniProt O75164; HPA: tissue IHC)UniProt lists a nuclear location. HPA tissue IHC reports cytoplasmic expression across tissues plus nuclear expression in neuronal and fallopian tube glandular cells. Record nuclear and cytoplasmic scores separately rather than forcing one consensus location (UniProt O75164; HPA: tissue IHC).
Strength of IHC evidence (HPA: reliability; HPA007610 validation)The tissue profile and HPA007610 IHC validation are Uncertain; HPA says external verification is pending. A matching image is therefore supportive of a working assay, while independent specificity controls remain important for interpretation (HPA: reliability; HPA007610 validation; standard IHC practice).
Protein form and epitope coverage (UniProt O75164)UniProt lists 2 isoforms, no signal peptide, and no transmembrane segment. The payload gives no antibody epitope, so it cannot establish which isoforms the IHC-validated antibody detects or predict an isoform-specific tissue pattern (UniProt O75164).
IF/ICC Q: Where should fluorescence localize? (HPA: subcellular ICC-IF)A: HPA reports a supported main location in the nucleoli fibrillar center and an additional supported location in the nucleoplasm. That ICC-IF observation informs image interpretation but does not establish the compartment or intensity of chromogenic tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
A reported High cell population is blank (HPA: tissue IHC).Possible assay or section failure; a blank section alone cannot identify the cause (standard IHC practice).Confirm the named cells are present, then review the positive control, antigen retrieval, and detection steps before scoring the result (HPA: tissue IHC; standard IHC practice).
Most of the section has diffuse brown color (standard IHC practice).Background may reflect nonspecific binding or endogenous detection activity; appearance alone cannot distinguish them (standard IHC practice).Inspect the negative detection control and blocking steps, then reassess whether staining is confined to identifiable cells and compartments (standard IHC practice).
Strong color appears in HPA Not detected cells (HPA: tissue IHC).Cross-reactivity or endogenous detection activity is possible; HPA's tissue profile is itself Uncertain (HPA: reliability; standard IHC practice).Compare the same run with a reported High population and negative detection control before assigning a KDM4A-positive score (HPA: tissue IHC; standard IHC practice).
Only cytoplasm stains, despite UniProt's nuclear assignment (UniProt O75164).HPA reports cytoplasmic tissue staining, so location alone cannot identify an artefact; its IHC profile remains Uncertain (HPA: tissue IHC; HPA: reliability).Record cytoplasmic and nuclear scores separately, verify morphology, and interpret the result with the assay controls (HPA: tissue IHC; standard IHC practice).
Only nuclei stain in a tissue expected to show cytoplasmic signal (HPA: tissue IHC).UniProt's nuclear assignment supports nuclear KDM4A, but it does not explain the missing HPA cytoplasmic pattern (UniProt O75164; HPA: tissue IHC).Check a reported High cell population in the same run and document the compartment discrepancy without assuming its cause (HPA: tissue IHC; standard IHC practice).
A proposed dilution or retrieval change is presented as KDM4A-specific (standard IHC practice).The supplied UniProt and HPA records provide no target-specific retrieval or fixation sensitivity data (UniProt O75164; HPA: tissue IHC).Treat such changes as assay optimization, compare controlled sections, and report the tested conditions without attributing an effect to KDM4A chemistry (standard IHC practice).

Sample controls for KDM4A IHC & IF

🧪Run duodenum first: glandular cells should stain (HPA: High in duodenal glandular cells). Use pancreas as a negative comparator (HPA: Not detected in pancreatic exocrine glandular cells); non-glandular cells on the duodenal slide should show only background if independently confirmed as internal negatives, since KDM4A is described as ubiquitous (UniProt: tissue specificity).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KDM4A in A-431, U-251MG, U2OS, with annotated localisation: Nucleoli fibrillar center (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a matched rabbit IgG isotype control (selected caption: rabbit primary and anti-rabbit IgG secondary), and KDM4A-knockout material as a biological negative if available (standard IHC practice). Quench endogenous peroxidase and check control slides for background when using HRP/DAB in duodenum (selected caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A KDM4A-specific fixation window or fixation effect is unreported; the selected-SKU paraffin-section caption does not report a fixative (selected caption: fixative not stated). Heat retrieval with EDTA at pH 8.0 was used, but whether that condition is required is unreported (selected caption: EDTA pH 8.0). The supplied evidence does not show that frozen sections or IF/ICC are easier; assess endogenous peroxidase background in duodenal HRP/DAB sections, and note that no KDM4A-specific tissue artefact is reported (selected caption: HRP/DAB; standard IHC practice).

HPA tissue IHC evidence for KDM4A

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. 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

TissueCell typeLevelEvidenceSource
Caudate Neuronal cells High Protein (IHC) HPA →
Cerebellum Cells in molecular layer High Protein (IHC) HPA →
Cerebral cortex Neuronal cells High Protein (IHC) HPA →
Duodenum Glandular cells High Protein (IHC) HPA →
Hippocampus Neuronal cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Pancreas Exocrine glandular cells Not detected Protein (IHC) HPA →
Salivary gland Glandular cells Not detected Protein (IHC) HPA →
Smooth muscle Smooth muscle cells Not detected Protein (IHC) HPA →
Soft tissue Peripheral nerve Not detected Protein (IHC) HPA →
Section 3

Advanced KDM4A IHC Tips

Troubleshoot chromogenic IHC for KDM4A in paraffin sections using the catalog antibody’s tissue staining conditions and compartment evidence (datasheet A02850-1; UniProt O75164).

What retrieval should I try first for weak KDM4A staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this paraffin-section assay (datasheet A02850-1). The documented tissue staining used 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions before changing retrieval (datasheet A02850-1). If staining remains weak, compare a modestly longer heating step with the original condition on adjacent sections, keeping cooling and detection consistent (standard IHC practice). Include a section processed without primary antibody to check whether stronger retrieval also increases background (standard IHC practice). Score nuclear signal separately because KDM4A is reported in the nucleus (UniProt O75164).
Could fixation explain weak or patchy KDM4A staining?
The KDM4A tissue-IHC caption identifies a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A02850-1). Record the fixative and fixation duration for each specimen, then compare sections processed with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A02850-1; standard IHC practice). Underfixation can impair tissue preservation, while excessive fixation can reduce access to some epitopes; these are general IHC possibilities, not established KDM4A effects (standard IHC practice). Use a well-preserved control section in each run and evaluate staining alongside morphology (standard IHC practice). Do not attribute a changed nuclear pattern to fixation without a controlled comparison (standard IHC practice).
How should I assess cytoplasmic staining when KDM4A is expected in nuclei?
Assess nuclear and cytoplasmic chromogen separately: UniProt places KDM4A in the nucleus, whereas HPA reports cytoplasmic tissue staining and nuclear staining in neuronal and fallopian-tube glandular cells (UniProt O75164; HPA tissue IHC). HPA’s subcellular IF data place signal mainly in the nucleoli fibrillar center, with additional nucleoplasmic signal (HPA subcellular). Because the HPA tissue-IHC reliability is uncertain, cytoplasmic staining alone needs independent validation before it is called KDM4A-specific (HPA tissue IHC). Compare staining with a no-primary control and inspect individual cells at high magnification (standard IHC practice). Report the fraction and intensity of each compartment rather than collapsing both into one score (standard IHC practice).
Could an isoform or altered epitope explain discordant staining?
KDM4A has 2 recorded isoforms, but the supplied tissue-IHC caption does not map the catalog antibody’s epitope to either one (UniProt O75164; datasheet A02850-1). Its JmjN domain spans residues 14–56, JmjC spans 142–308, and Tudor domains span 897–1011; those positions cannot establish antibody coverage without an epitope sequence (UniProt O75164). The record also lists acetylation at residue 2 and phosphorylation at residue 523, without evidence that either changes IHC binding (UniProt O75164). Request the immunogen or epitope information and check its presence in each isoform before assigning a staining difference to splicing (standard IHC practice). Keep retrieval and detection matched while comparing specimens (standard IHC practice).
How can I adapt the KDM4A question to multiplex IF?
For IF, pair KDM4A with an independently validated marker for the cell population being assessed, such as neuronal cells when examining the HPA-reported nuclear pattern (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue, placing the weaker target in a channel with low autofluorescence and minimal overlap with the companion marker (standard IF practice). Permeabilise sufficiently to access nuclear antigen because KDM4A is nuclear and has no transmembrane segment, then compare permeabilised and minimally permeabilised controls (UniProt O75164; standard IF practice). HPA IF observations include nucleoli fibrillar-center and nucleoplasmic localisation, but they do not establish an IF protocol for this catalog antibody (HPA subcellular; datasheet A02850-1). Acquire single-label controls before interpreting colocalisation (standard IF practice).
How do I reduce diffuse DAB staining without losing nuclear signal?
Begin with the documented 10% goat-serum block and 2 μg/ml primary antibody, incubated overnight at 4°C, as a reference condition (datasheet A02850-1). Include a no-primary section to assess secondary reagent and chromogen background, and block endogenous peroxidase before HRP–DAB detection (standard IHC practice). If background persists, titrate the primary antibody downward and adjust wash duration while keeping retrieval at EDTA pH 8.0 for the initial comparison (datasheet A02850-1; standard IHC practice). Examine tissue edges and damaged areas separately because they can stain nonspecifically (standard IHC practice). Preserve any reproducible nuclear signal during optimisation, given KDM4A’s nuclear localisation (UniProt O75164).
What scoring method makes KDM4A IHC results comparable across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine the cell population and score nuclear and cytoplasmic staining separately because the localisation evidence differs between UniProt and HPA tissue IHC (UniProt O75164; HPA tissue IHC). For each compartment, report the percentage of positive cells and an H-score calculated as 1 × % weak + 2 × % moderate + 3 × % strong (standard IHC practice). Count within a fixed viable tissue area or report positive cells per mm², excluding folds and necrosis (standard IHC practice). Normalise counts to the number of evaluable cells in the same compartment and tissue region, using consistent exposure to chromogen and counterstain (standard IHC practice). Document scoring thresholds before comparing groups (standard IHC practice).
Which staining patterns warrant caution before calling KDM4A positive?
A reproducible nuclear signal is compatible with KDM4A’s annotated localisation, while HPA tissue IHC also reports cytoplasmic staining with uncertain reliability (UniProt O75164; HPA tissue IHC). Check whether positive cells match the tissue context: HPA reports high staining in neuronal cells of the caudate and no detection in adipocytes, though these observations are not definitive controls (HPA tissue IHC). Treat strong edge staining, necrotic areas and signal on a no-primary slide as possible artefacts (standard IHC practice). For HRP–DAB assays, assess endogenous peroxidase with an appropriate control before assigning diffuse brown signal to KDM4A (standard IHC practice). Confirm disputed compartment patterns with an independent antibody or orthogonal method (standard IHC practice).
Boster reagents

Best KDM4A / Lysine-specific demethylase 4A IHC Antibodies

Two anti-KDM4A antibodies have real IHC images covering human and mouse paraffin sections and rat liver tissue (A02850-1 and A02850 image captions); no IF images are supplied (catalog: IF image alts).

Real IHC data IHC analysis of JHDM3A/JMJD2A/KDM4A using anti-JHDM3A/JMJD2A/KDM4A antibody (A02850-1). JHDM3A/JMJD2A/KDM4A was detected in a paraffin-embedded section of human ovarian serous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-JHDM3A/JMJD2A/KDM4A Antibody (A02850-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-JHDM3A/JMJD2A/KDM4A Antibody ®
Cat # A02850-1
Real IHC data Immunohistochemistry of JMJD2A in rat liver tissue with JMJD2A antibody at 5 μg/mL.
Anti-JMJD2A KDM4A Antibody
Cat # A02850

A02850-1 has IHC images from human ovarian serous carcinoma and mouse colon paraffin sections (A02850-1 image captions). A02850 has a rat liver IHC image at 5 μg/mL (A02850 image caption).

Which to pick: For cross-species paraffin-section IHC in human and mouse samples, choose A02850-1: its own captions show both species using 2 μg/ml antibody after EDTA heat retrieval (A02850-1 image captions). For rat tissue IHC, choose A02850, whose own image shows rat liver at 5 μg/mL; its listed human and mouse reactivity does not establish IHC validation in those species (A02850 image caption; catalog: reactivity). Neither SKU has documented IF/ICC use or IF images, and the fixative is unreported in both IHC captions (catalog: applications and IF image alts; A02850-1 and A02850 image captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry O75164 (KDM4A_HUMAN, Lysine-specific demethylase 4A).
  2. Human Protein Atlas. KDM4A tissue IHC expression (reliability: Uncertain).
  3. Human Protein Atlas. KDM4A subcellular location (ICC-IF): Mainly localized to the nucleoli fibrillar center. In addition localized to the nucleoplasm..
  4. Human Protein Atlas. KDM4A antibody validation summary (1 antibodies).
  5. KDM4A-driven SMAD5/TGFβ axis activation as a therapeutic target in cholangiocarcinoma: mechanistic insights and translational implications. Cell & bioscience 2025 — PMC12482258.
  6. KDM4B and KDM4A promote endometrial cancer progression by regulating androgen receptor, c-myc, and p27kip1. Oncotarget 2015 — PMC4741634.
  7. Histone demethylase KDM4A plays an oncogenic role in nasopharyngeal carcinoma by promoting cell migration and invasion. Experimental & molecular medicine 2021 — PMC8417295.
  8. Targeting USP1-dependent KDM4A protein stability as a potential prostate cancer therapy. Cancer science 2020 — PMC7226285.
  9. PubMed PMID:9734811 — UniProt-cited evidence.
  10. PubMed PMID:16710414 — UniProt-cited evidence.
  11. PubMed PMID:15489334 — UniProt-cited evidence.