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KAT2B / Histone acetyltransferase KAT2B · IHC design guide
Design Immunohistochemistry for KAT2B
Plan KAT2B IHC in paraffin sections around the general nuclear staining reported in tissues (HPA tissue IHC). This guide helps select a high-staining control, handle fixation consistently and interpret differentiation-related changes in localisation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KAT2B 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 | General nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining; high in placental decidual cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A02403-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02403-1) | |
| Caveat | Differentiation can shift staining from cytoplasm to nucleus (UniProt) | |
| Regulation | Nuclear signal varies with differentiation (UniProt) | |
| Isoform / epitope | No annotated isoforms; one unprocessed chain (UniProt) |
Section 1
Recommended KAT2B IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses Tris-EDTA pH 9.0 retrieval (datasheet A02403-1). Published KAT2B IHC methods provide cervical and nasopharyngeal carcinoma examples (PMC8806817; PMC7719841).
| Sample | Paraffin-embedded human cervical carcinoma tissue; fixative not specified (datasheet A02403-1) |
| Fixation | Image fixative and duration unreported (datasheet A02403-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A02403-1); 20 min, 95–100 °C (standard) |
| 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-KAT2B, 1:50 recommended; image 1:200 (datasheet A02403-1) |
| 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 | KAT2B-positive staining in cells in molecular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 heat retrieval for the catalog antibody (datasheet A02403-1); the cervical carcinoma protocol used heated citrate buffer (PMC8806817).
Section 2
What Is the Expected KAT2B Staining Pattern?
KAT2B should stain mainly nuclei in paraffin sections (UniProt Q92831; HPA tissue IHC: general nuclear expression). HPA reports High staining in selected cell populations, including salivary glandular cells, placental decidual cells, and skin melanocytes (HPA tissue IHC). Tissue IHC reliability is Supported, with medium consistency against RNA data (HPA tissue IHC). KAT2B has no transmembrane segment (UniProt Q92831 topology).
| Distinct nuclear chromogen in salivary glandular cells, placental decidual cells, or skin melanocytes (HPA tissue IHC: High). | This fits the reported cell populations and general nuclear pattern (HPA tissue IHC). Compare staining with nuclear morphology and a counterstain when judging individual cells (general IHC practice). |
| Predominantly diffuse cytoplasmic staining with little nuclear signal across a section. | Review specificity and detection background because the reported tissue pattern is generally nuclear (HPA tissue IHC). Cytoplasmic staining alone is not conclusive: localization can vary with differentiation (UniProt Q92831). |
| Strong chromogen in a cell population outside the reported High populations, especially if its nuclei remain unstained. | Consider cross-reactivity or endogenous detection activity and inspect controls (general IHC practice). HPA lists no negative tissue, so an unlisted cell population cannot be classified as KAT2B-negative (HPA tissue IHC). |
| Diffuse color over cells, stroma, and spaces, without clear nuclear boundaries. | This is difficult to score as the general nuclear pattern reported for KAT2B (HPA tissue IHC). Check the no-primary control and detection background before assigning cell-specific expression (general IHC practice). |
| No discernible signal in an HPA High population, such as salivary glandular cells (HPA tissue IHC). | Treat the result as inconclusive until controls and the IHC-validated antibody are checked (general IHC practice). HPA's Supported tissue assessment indicates medium consistency, not guaranteed staining in every section (HPA tissue IHC). |
💡Expected KAT2B appearanceA convincing positive has predominantly nuclear staining in the reported cell population, with High signal in selected HPA populations; widespread compartment-free color is suspect background (HPA tissue IHC; general IHC practice).
| Cell and tissue choice | HPA reports High staining in cerebellar molecular-layer cells, decidual cells, salivary and stomach glandular cells, melanocytes, thyroid glandular cells, and urothelial cells; bone-marrow hematopoietic cells are Medium (HPA tissue IHC). Choose a documented High population for interpretation. |
| Compartment | Mainly nuclear localization is supported by UniProt and tissue IHC (UniProt Q92831; HPA tissue IHC). UniProt also records cytoplasm and centrosomes, so a limited nonnuclear component needs cell-context review rather than automatic rejection (UniProt Q92831). |
| Differentiation state | UniProt reports cytoplasmic KAT2B in undifferentiated basal epithelial cells and nuclear KAT2B in differentiated parabasal cells (UniProt Q92831). Assess the layer and cell state before treating a compartment shift as staining failure. |
| Antibody evidence | CAB004526 has an IHC Supported designation; the listed HPA antibodies with ICC Enhanced designations have no IHC designation in this payload (HPA antibody validation). Use the IHC evidence when interpreting paraffin-section chromogen. |
| Target structure | KAT2B is a single listed chain with no signal peptide, propeptide, or transmembrane segment (UniProt Q92831). The supplied record gives no antibody epitope or target-specific fixation effect, so neither epitope access nor fixation sensitivity can be predicted from it. |
| IF/ICC Q: what pattern is expected? | A: Mainly nucleoplasmic signal with additional cytosol signal is reported in cell images (HPA ICC-IF). That observation helps interpret compartment differences; the IF/ICC workflow belongs on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a documented High cell population (HPA tissue IHC). | The section, antibody use, or detection workflow may have failed; absence of color alone cannot identify which step failed (general IHC practice). | Check a documented positive section, the IHC-validated antibody's stated conditions, reagent performance, and the no-primary control before interpreting absence as biological (general IHC practice). |
| Diffuse chromogen obscures nuclei. | Nonspecific primary staining or detection background can reduce contrast (general IHC practice). | Compare a no-primary control, review blocking and washing, and optimize primary-antibody concentration within its stated IHC conditions (general IHC practice). |
| Cytoplasmic color dominates where a general nuclear pattern was expected (HPA tissue IHC). | Cell state may contribute, since KAT2B localization varies with differentiation; technical background is another possibility (UniProt Q92831; general IHC practice). | Check cell identity and layer, then compare nuclear detail and controls. Interpret a compartment shift in that context rather than calling it positive solely from color (UniProt Q92831; general IHC practice). |
| Unexpected cells stain strongly while reported High cells are weak. | Cross-reactivity or endogenous detection activity is possible; HPA provides no negative tissue reference here (general IHC practice; HPA tissue IHC). | Compare the no-primary control and a documented High cell population in the same run; avoid calling an unlisted population negative by definition (general IHC practice; HPA tissue IHC). |
| A low-staining tissue shows faint nuclear color. | Low HPA staining is not an absence claim; liver hepatocytes and several other listed cells have Low staining (HPA tissue IHC). | Score intensity and cellular localization against controls, and report the observation without imposing a positive-versus-negative cutoff unsupported by this payload (HPA tissue IHC; general IHC practice). |
| Color persists in the no-primary control. | Endogenous enzyme activity or detection-reagent background can produce chromogen independently of primary-antibody binding (general chromogenic IHC practice). | Address the detection system's background according to its instructions and repeat the control before attributing sample color to KAT2B (general chromogenic IHC practice). |
Experiment evidence
IHC & IF images showing KAT2B staining
A02403-1 · IHCImmunohistochemical analysis of paraffin-embedded human cervical carcinoma. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, p
A02403 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using p300/CBP Antibody. The picture on the right is block
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114166-KAT2B/tissue/cerebral+cortex#imid_1604516
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114166-KAT2B/tissue/colon#imid_1604450
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114166-KAT2B/tissue/liver#imid_1604435
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000114166-KAT2B/tissue/kidney#imid_1604436
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA002470; green = KAT2B, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA002470; green = KAT2B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KAT2B IHC & IF
🧪Run salivary gland first and assess staining in its glandular cells (HPA: High in salivary gland glandular cells). HPA detects KAT2B in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells without visible signal on the positive slide as background comparators rather than proven biological negatives (HPA: no negative tissue rows; standard IHC practice).
Positive control tissue: Cerebellum (Cells in molecular layer, HPA High)
Negative control tissue: None in HPA: KAT2B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KAT2B in HEL, SK-MEL-30, U2OS, HeLa, U-251MG, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype control matched to the primary antibody’s host species and clonality; a KAT2B knockout sample provides a biological specificity control (standard IHC practice). For salivary gland IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU A02403-1 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption uses Tris–EDTA at pH 9.0 for retrieval, but does not establish that KAT2B staining depends on this retrieval condition (selected-SKU tissue-IHC caption). Frozen-section performance and whether IF/ICC is easier are unreported; salivary gland glandular staining should be judged against any luminal or nonspecific chromogen background (HPA: High in salivary gland glandular cells; standard IHC practice).
HPA tissue IHC evidence for KAT2B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Skin | Melanocytes | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: KAT2B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced KAT2B IHC Tips
Troubleshoot KAT2B staining in paraffin sections by checking retrieval, nuclear localisation, cell type and controls before interpreting chromogenic signal.
What retrieval should I try first if KAT2B staining is weak?
Start with heat-mediated Tris-EDTA retrieval at pH 9.0 for paraffin sections (datasheet A02403-1). The selected image used that buffer with a 1:200 primary antibody incubation overnight at 4 °C, providing a starting condition for comparison (caption A02403-1). If staining remains weak, vary heating duration or try a different retrieval buffer as a controlled fallback, processing a reference section alongside each change (standard IHC practice). Judge improvement by clearer nuclear staining and preserved tissue morphology, since KAT2B is mainly nuclear and HPA reports general nuclear tissue expression (UniProt Q92831 localisation; HPA tissue IHC profile).
Could fixation explain weak or uneven KAT2B staining?
The selected paraffin-section caption does not state a fixative, so KAT2B-specific fixation sensitivity is unknown (caption A02403-1). Record the fixative, fixation duration and specimen thickness for each section, then compare sections processed under matched conditions before changing antibody concentration (standard IHC practice). Excessive or uneven fixation can alter antigen accessibility in IHC generally, but these payloads do not establish that effect for KAT2B (standard IHC practice; caption A02403-1). Run the documented pH 9.0 Tris-EDTA retrieval consistently, and assess morphology and nuclear staining together when evaluating a fixation change (datasheet A02403-1; UniProt Q92831 localisation).
Should cytoplasmic or punctate staining count as KAT2B positive?
Expect predominantly nuclear chromogenic staining because KAT2B mainly localises to the nucleus and HPA describes general nuclear expression in tissue (UniProt Q92831 localisation; HPA tissue IHC profile). Cytoplasmic staining can be biologically plausible in some differentiation states; UniProt reports cytoplasmic basal epithelial cells and nuclear parabasal cells (UniProt Q92831 localisation). Centrosomal localisation around late G1 and the G1/S transition could produce small extranuclear foci, but diffuse cytoplasmic colour alone cannot establish that identity in a section (UniProt Q92831 localisation; standard IHC interpretation). Score nuclear and cytoplasmic compartments separately, and compare their patterns with tissue architecture and negative controls (standard IHC practice).
How can I investigate staining that changes across differentiation states?
The supplied record annotates 0 isoforms, while the selected caption does not identify the antibody epitope; do not assign a staining difference to a specific KAT2B isoform or epitope (UniProt Q92831 record; caption A02403-1). KAT2B has an N-acetyltransferase domain at residues 503–651 and a bromodomain at 723–827, which describe protein regions rather than the antibody's binding site (UniProt Q92831 domains). Request epitope information or compare an independently validated antibody when specificity remains uncertain (standard IHC practice). Compare basal and parabasal compartments separately because their reported cytoplasmic-to-nuclear shift could affect a nuclear-only score (UniProt Q92831 localisation).
How should IF help assess an ambiguous KAT2B IHC pattern?
Use IF as a complementary localisation check, since HPA reports enhanced nucleoplasmic and additional cytosolic localisation in its subcellular data (HPA subcellular profile). Multiplex KAT2B with a validated marker of the expected cell type, such as melanocytes when evaluating skin, and include a nuclear counterstain to assign signal to individual cells (HPA tissue IHC: high in melanocytes; standard IF practice). Choose a far-red fluorophore when tissue autofluorescence compromises shorter wavelengths, checking an unstained section in the same channels (standard IF practice). Because KAT2B has no transmembrane segment and is mainly intracellular, optimise permeabilisation to expose nuclear antigen while preserving cell boundaries (UniProt Q92831 topology and localisation; standard IF practice).
What should I check when the entire section looks brown?
First compare a no-primary control with the stained section to identify detection-system background, and inspect whether colour follows tissue edges or damaged areas (standard chromogenic IHC practice). Quench endogenous peroxidase before an HRP/DAB reaction and check that blocking, washes and secondary antibody conditions suit the tissue and detection system (standard chromogenic IHC practice). The selected caption reports a 1:200 secondary antibody for 45 minutes at room temperature; treat this as an image condition, not proof that it is optimal for every section (caption A02403-1). Titrate the primary around its documented 1:200 condition while retaining a reference section and a no-primary control (caption A02403-1; standard IHC practice).
How should I score KAT2B across mixed cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear compartment before scoring, because HPA reports general nuclear expression while UniProt describes differentiation-dependent shifts between cytoplasm and nucleus (HPA tissue IHC profile; UniProt Q92831 localisation). Record the percentage of positive nuclei and staining intensity, then calculate a nuclear H-score from 0–300 if an ordinal intensity scale is appropriate (standard IHC scoring practice). Normalise positive counts to all evaluable nuclei in the same annotated cell population, or report positive-cell density per mm² of evaluable tissue when area is the denominator (standard IHC scoring practice). Exclude folds, necrosis and section edges consistently, and document the threshold using matched controls (standard IHC practice).
What evidence separates genuine KAT2B signal from artefact?
A convincing result shows reproducible staining in the expected cells with predominantly nuclear localisation, consistent with HPA's general nuclear tissue profile and UniProt's main nuclear localisation (HPA tissue IHC profile; UniProt Q92831 localisation). For context, HPA reports high staining in salivary-gland glandular cells and low staining in liver hepatocytes, but its tissue-IHC reliability is only Supported with medium RNA concordance (HPA tissue IHC). Treat colour confined to edges, necrosis or a no-primary control as possible processing or endogenous-enzyme artefact, especially in chromogenic detection (standard IHC practice). Investigate cytoplasmic signal against cell differentiation and controls before rejecting it, because KAT2B localisation can change with differentiation (UniProt Q92831 localisation).
Boster reagents
Best KAT2B / Histone acetyltransferase KAT2B IHC Antibodies
Catalog antibodies cover IHC in human paraffin carcinoma sections (A02403 and A02403-1 image captions), with human, mouse, and rat reactivity across the range (catalog). IF is listed, but no IF figure is supplied (catalog).
Real IHC data
Anti-Acetyl-PCAF (K428) KAT2B Antibody
Cat # A02403-1
Real IHC data
Anti-PCAF KAT2B Antibody
Cat # A02403
A02403-1 has an IHC image from paraffin-embedded human cervical carcinoma and lists human and mouse reactivity (A02403-1 image caption; catalog). A02403 has a paraffin-embedded human breast carcinoma IHC caption that names p300/CBP, while A02403-3 lists human IHC, IF, and ICC without an image (A02403 image caption; A02403-3 catalog).
Which to pick: For tissue IHC targeting acetyl-K428, choose A02403-1: its own paraffin-section caption reports Tris-EDTA pH 9.0 retrieval and 1:200 antibody overnight at 4 °C; the fixative is unreported (A02403-1 catalog; image caption). For human IF/ICC, A02403-3 lists both applications at 5 μg/ml, but supplies no IF image (A02403-3 catalog). For cross-species IHC or IF, A02403 lists human, mouse, and rat reactivity and both applications; its breast carcinoma caption names p300/CBP, so verify target identity before use, and note that the fixative is unreported (A02403 catalog; image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92831 (KAT2B_HUMAN, Histone acetyltransferase KAT2B).
- Human Protein Atlas. KAT2B tissue IHC expression (reliability: Supported).
- Human Protein Atlas. KAT2B subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. KAT2B antibody validation summary (4 antibodies).
- Lysine Acetyltransferase 2B predicts favorable prognosis and functions as anti-oncogene in cervical carcinoma. Bioengineered 2021 — PMC8806817.
- Upregulation of cGMP-dependent Protein Kinase (PRKG1) in the Development of Adolescent Idiopathic Scoliosis. Orthopaedic surgery 2020 — PMC7454216.
- KAT2B inhibits proliferation and invasion via inactivating TGF-β/Smad3 pathway-medicated autophagy and EMT in epithelial ovarian cancer. Scientific reports 2025 — PMC11772695.
- Long Noncoding RNA AFAP1-AS1 Is a Critical Regulator of Nasopharyngeal Carcinoma Tumorigenicity. Frontiers in oncology 2020 — PMC7719841.
- PubMed PMID:8684459 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:8945521 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KAT2B (Histone acetyltransferase KAT2B). Expected localisation General nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A02403-1). Validated IHC antibody: A02403-1 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in molecular layer); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human cervical carcinoma tissue; fixative not specified (datasheet A02403-1); 2. Fixation: Image fixative and duration unreported (datasheet A02403-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Tris-EDTA pH 9.0 (datasheet A02403-1); 20 min, 95–100 °C (standard); 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-KAT2B, 1:50 recommended; image 1:200 (datasheet A02403-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02403-1); Caveat: Differentiation can shift staining from cytoplasm to nucleus (UniProt); Isoform / epitope: No annotated isoforms ; one unprocessed chain (UniProt); Regulation: Nuclear signal varies with differentiation (UniProt). Evidence: UniProt Q92831, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A02403-1, PMC8806817, PMC7719841. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KAT2B/immunohistochemistry-ihc-guide.