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HAT1 / Histone acetyltransferase type B catalytic subunit · IHC design guide
Design Immunohistochemistry for HAT1
Use colon glandular cells as a positive reference and adipocytes as a negative reference for the nuclear staining seen in most tissues (HPA tissue IHC). Catalog antibody M03596 has a documented paraffin IHC example using 1:50 overnight at 4°C with HRP/DAB detection (datasheet M03596).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HAT1 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 | Predominantly nuclear in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03596) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Oxidative stress enhances nuclear localisation (UniProt) | |
| Isoform / epitope | 2 isoforms (A/B); epitope impact unknown (UniProt) |
Section 1
Recommended HAT1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet M03596) with published HAT1 staining methods for myocardium (PMC9715834) and mouse ovary (PMC10617186).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet M03596) |
| Fixation | Image fixative and duration unreported (datasheet M03596); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M03596); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03596) |
| Primary antibody | Rabbit monoclonal (clone 21H24) anti-HAT1, 1:50 (datasheet M03596) |
| Primary incubation | Overnight at 4 °C (datasheet M03596) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03596) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HAT1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet M03596); the mouse ovary protocol uses Tris-EDTA pH 9.0 (PMC10617186).
Section 2
What Is the Expected HAT1 Staining Pattern?
Expect predominantly nuclear HAT1 staining in many cell types, including glandular cells of the colon and hematopoietic cells of bone marrow (HPA tissue IHC: nuclear expression in most tissues; High in those cells). Nucleoplasmic staining is consistent with the subcellular record (HPA ICC-IF: nucleoplasm supported). The tissue IHC profile has Supported reliability with medium consistency against RNA data (HPA tissue IHC). HAT1 has no transmembrane segment (UniProt O14929 topology).
| Clear nuclear chromogen in colon glandular cells or bone marrow hematopoietic cells, with nuclei distinguishable from the counterstain. | This fits the reported high staining in those cells and the broader nuclear tissue profile (HPA tissue IHC: High in colon glandular cells and bone marrow hematopoietic cells; nuclear expression in most tissues). Compare signal within the relevant cell population rather than judging the section by its overall darkness (general IHC practice). |
| Predominantly cytoplasmic or membrane-like staining with little nuclear signal. | Treat this as discordant with the expected pattern and investigate an artefact or nonspecific detection (HPA tissue IHC: nuclear expression in most tissues; UniProt O14929: predominantly nuclear in normal cells, no transmembrane segment). UniProt also lists cytoplasm and mitochondrion, so a secondary signal alone does not establish an artefact (UniProt O14929 subcellular location). |
| Strong staining in adipocytes or lung alveolar cells while the expected nuclear pattern is absent from known-positive cells. | Check for cross-reactivity or endogenous detection activity (general IHC practice). These particular cell populations were reported as Not detected, while colon glandular cells were High (HPA tissue IHC). A negative call for one cell type does not make its entire tissue a negative control (HPA tissue IHC: cell-specific entries). |
| Diffuse brown haze over nuclei, cytoplasm, and extracellular areas, including the negative-control section. | The distribution cannot be scored as HAT1-specific nuclear staining; consider residual detection background, inadequate blocking, or excessive reagent concentration (general IHC practice). The reference pattern is nuclear in most tissues (HPA tissue IHC). Compare an appropriately matched negative control before assigning a cellular score (general IHC practice). |
| No nuclear signal in colon glandular cells or bone marrow hematopoietic cells. | These are useful positive comparators because both were High in the supplied tissue profile (HPA tissue IHC). Check that the expected cells are present, then review staining and detection controls (general IHC practice). A failed control makes a negative result in the test section uninterpretable (general IHC practice). |
💡Expected HAT1 appearanceCall a convincing positive result when distinct, cell-associated nuclear chromogen is present in glandular or hematopoietic cells reported as High (HPA tissue IHC: colon glandular cells; bone marrow hematopoietic cells); broad cytoplasmic haze or staining outside recognizable cells is a suspect positive (general IHC practice).
| Cell-type selection for IHC controls | Colon glandular cells and bone marrow hematopoietic cells were High; adipocytes and lung alveolar cells were Not detected (HPA tissue IHC). Score the named cells, since the entries do not classify every cell in those tissues (HPA tissue IHC: cell-specific profile). |
| Subcellular localization and biological context | HAT1 is predominantly nuclear in normal cells; hydrogen peroxide or ionizing radiation enhances nuclear localization by redistributing existing protein (UniProt O14929 subcellular location). Record treatment context when comparing compartments. |
| Antibody evidence | Two listed antibodies have Supported IHC status, and HPA036788 also has Supported ICC status (HPA antibodies: HPA036788, CAB037191). The tissue profile itself is Supported with medium consistency against RNA data; that rating is not proof that every stained cell is specific (HPA tissue IHC). |
| Isoforms and processing | Two isoforms, A and B, are listed, along with a chain spanning residues 2–419 (UniProt O14929 isoforms; processing). No epitope position is supplied, so these annotations cannot predict which isoform the catalog antibody detects or a distinct IHC pattern. |
| IF/ICC Q&A: What localization should be expected? | Nucleoplasmic signal is the supported IF/ICC location (HPA ICC-IF: Nucleoplasm supported). The listed image cell lines are A-431, U-251MG, and U2OS (HPA ICC-IF); their listing does not supply an IHC tissue result or an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The test section is blank and the colon positive comparator is also blank. | A shared staining or detection failure is possible; colon glandular cells were High in the reference profile (HPA tissue IHC). | Confirm glandular cells are present, then inspect primary-antibody application, detection reagents, and control performance before interpreting the test section (general IHC practice). |
| Signal is diffuse across cells and the negative control. | Background from blocking or the detection system is possible (general IHC practice). | Review the negative control and blocking steps; adjust reagent concentration or wash conditions using the chosen IHC workflow (general IHC practice). Reassess whether discrete nuclear staining emerges (HPA tissue IHC: nuclear profile). |
| Brown signal is strongest outside nuclei. | Nonspecific staining or compartment misidentification is possible (general IHC practice); predominant nuclear localization is expected in normal cells (UniProt O14929 subcellular location). | Use the counterstain to locate nuclei and compare a positive tissue control. Record any oxidative or radiation treatment before interpreting a compartment shift (general IHC practice; UniProt O14929 subcellular location). |
| Adipocytes or lung alveolar cells appear strongly positive. | Those named cells were Not detected in the HPA tissue observations, so cross-reactivity or endogenous detection activity warrants investigation (HPA tissue IHC; general IHC practice). | Check cell identification and matched negative controls, then compare nuclear staining in a reported high cell population before assigning specificity (HPA tissue IHC: colon glandular cells High; general IHC practice). |
| Nuclear staining varies greatly between sampled tissues or cell populations. | The HPA entries differ by cell type, and the tissue profile has medium consistency with RNA data (HPA tissue IHC). A whole-section average can hide the named positive population (general IHC practice). | Score comparable cell populations separately and document tissue, cell type, and nuclear intensity; keep the HPA cell-specific observations as the reference (HPA tissue IHC; general IHC practice). |
| A change in antigen retrieval is proposed to explain weak HAT1 staining. | Target-specific fixation or retrieval sensitivity is unreported in the supplied UniProt and HPA records. | Treat retrieval adjustment as general IHC optimization, with a known-positive tissue and matched controls in each comparison (general IHC practice; HPA tissue IHC: colon glandular cells High). Do not infer an HAT1-specific fixation effect from the staining profile. |
Experiment evidence
IHC & IF images showing HAT1 staining
M03596 · IHCIHC analysis of KAT1 using anti-KAT1 antibody (M03596). KAT1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat
M03596 · IHCIHC analysis of KAT1 using anti-KAT1 antibody (M03596). KAT1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat
A03596-2 · IHCIHC analysis of KAT1/HAT1 using anti-KAT1/HAT1 antibody (A03596-2). KAT1/HAT1 was detected in a paraffin-embedded section of human liver can
A03596-2 · IHCIHC analysis of KAT1/HAT1 using anti-KAT1/HAT1 antibody (A03596-2). KAT1/HAT1 was detected in a paraffin-embedded section of human lung canc
A03596-2 · IFIF analysis of KAT1/HAT1 using anti-KAT1/HAT1 antibody (A03596-2) and anti-Tubulin Alpha antibody (M03989-3). KAT1/HAT1 was detected in immu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000128708-HAT1/tissue/cerebral+cortex#imid_9302184
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000128708-HAT1/tissue/colon#imid_9300822
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000128708-HAT1/tissue/liver#imid_9300823
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000128708-HAT1/tissue/kidney#imid_9300816
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036788; green = HAT1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036788; green = HAT1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HAT1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the colon slide, cells without nuclear staining can show background, but should not be assumed to be HAT1-negative without validation.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HAT1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) section, a host- and clonality-matched rabbit IgG isotype control, and HAT1-knockout material as a biological negative (selected-SKU caption: rabbit primary antibody). For chromogenic detection in colon, quench endogenous peroxidase and check for tissue background before scoring DAB (selected-SKU caption: HRP/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the fixative for M03596 is unreported (selected-SKU caption: paraffin section; fixative not stated). Start IHC retrieval optimization from heat-mediated EDTA at pH 8.0, as used in the selected-SKU caption, but its necessity for other specimens is unreported. The evidence does not establish that frozen sections or IF are easier; for IF/ICC, nucleoplasmic signal is the supported localization to assess (HPA: Nucleoplasm supported).
HPA tissue IHC evidence for HAT1
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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HAT1 IHC Tips
Troubleshoot HAT1 staining in paraffin sections by assessing retrieval, nuclear localisation, controls and scoring together (datasheet M03596; HPA tissue IHC).
What should I change when HAT1 staining is weak after antigen retrieval?
Use heat-mediated EDTA at pH 8.0 for HAT1 retrieval in paraffin sections (datasheet M03596). The selected image used that retrieval, followed by 10% goat serum blocking and 1:50 primary antibody overnight at 4°C (datasheet M03596). If nuclear staining remains weak, compare retrieval heating and cooling conditions on matched sections before changing the primary incubation; inspect tissue integrity and include a no-primary control (standard IHC practice). Record each condition because nuclear expression is expected in most tissues, but its intensity varies by cell population (HPA tissue IHC).
Could fixation explain faint or uneven nuclear HAT1 staining?
The paraffin-section image does not state a fixative, so HAT1-specific fixation sensitivity is unknown (datasheet M03596). Record fixation type and duration for each specimen, and compare matched sections using heat-mediated EDTA at pH 8.0 before changing antibody concentration (standard IHC practice; datasheet M03596). If nuclei remain faint, assess morphology and a no-primary control while testing fixation and retrieval conditions with a staining-positive specimen (standard IHC practice). The reported nuclear tissue pattern and HAT1 modification sites do not establish that any particular fixative masks this antibody’s epitope (HPA tissue IHC; UniProt O14929).
How should I assess cytoplasmic HAT1 staining in a paraffin section?
Assess nuclear staining first: HAT1 is predominantly nuclear in normal cells, and the tissue profile reports nuclear expression in most tissues (UniProt O14929; HPA tissue IHC). Record cytoplasmic staining separately because cytoplasm and mitochondrion are also annotated locations, while the supported cell imaging location is nucleoplasm (UniProt O14929; HPA subcellular). Check whether the same cell population shows reproducible staining across intact section interiors and whether no-primary controls remain clear (standard IHC practice). Document treatment history when available, since hydrogen peroxide or ionizing radiation can enhance nuclear localisation through redistribution of existing HAT1 (UniProt O14929).
Can this IHC signal distinguish HAT1 isoforms or modification states?
Do not assign a paraffin-section signal to isoform A or B without documented antibody epitope coverage and isoform-specific validation (UniProt O14929; standard IHC interpretation). HAT1 has 2 annotated isoforms, but the selected image supplies neither an epitope sequence nor isoform discrimination data (UniProt O14929; datasheet M03596). Likewise, annotated acetylation and phosphorylation sites, including phosphoserine at residues 190 and 343, do not show whether this antibody detects modified protein differently (UniProt O14929). Compare the antibody documentation with isoform sequences and use appropriately validated controls before making an isoform or modification claim (standard IHC practice).
How should I plan a parallel IF check of HAT1 localisation?
For a parallel IF/ICC experiment, pair HAT1 with a validated marker of the cell population being assessed and a nuclear counterstain (standard IF practice; HPA tissue IHC). Choose fluorophores after checking specimen autofluorescence, favouring a spectrally separated longer-wavelength channel when background is strongest at shorter wavelengths (standard IF practice). Plan permeabilisation for access to an intracellular, predominantly nuclear target; HAT1 has no annotated transmembrane segment, while this antibody’s epitope location is unspecified (UniProt O14929; datasheet M03596). Include single-channel and no-primary controls, and validate IF conditions independently of the paraffin-section 1:50 IHC incubation (standard IF practice; datasheet M03596).
What causes diffuse brown staining despite an apparently positive HAT1 control?
Compare the stained section with a no-primary control to assess detection-system and tissue background before interpreting diffuse DAB colour as HAT1 (standard IHC practice). Apply a peroxidase block appropriate to the specimen and detection system; this is a general chromogenic IHC step, while the selected image documents peroxidase detection and DAB development (standard IHC practice; datasheet M03596). Check whether excess primary antibody, incomplete blocking or section drying explains background, then adjust one variable at a time against the documented 10% goat serum and 1:50 primary conditions (standard IHC practice; datasheet M03596). Interpret convincing signal chiefly in intact cell nuclei and compare cell populations with reported HAT1 staining patterns (HPA tissue IHC; standard IHC practice).
How can I quantify HAT1 staining across heterogeneous paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, then report nuclear percent-positive cells and intensity using a consistent threshold (standard IHC scoring practice; HPA tissue IHC). An H-score combines the percentages at intensities 0–3 into a 0–300 score; positive nuclear density per mm² can supplement it when cell numbers differ (standard IHC scoring practice). Normalise positive counts to all eligible nuclei in the same compartment, and report area separately if comparing densities (standard IHC scoring practice). Keep retrieval, detection, imaging and scoring settings matched because the tissue profile includes high, low and undetected cell populations (datasheet M03596; HPA tissue IHC; standard IHC practice).
When is apparent HAT1 positivity more likely to be artefact?
Seek reproducible nuclear staining in intact cells, consistent with predominantly nuclear HAT1 and the reported tissue pattern (UniProt O14929; HPA tissue IHC). Compare an expected higher-staining population, such as appendix glandular cells, with a reported undetected population, such as adipocytes; neither alone proves antibody specificity (HPA tissue IHC; standard IHC interpretation). Treat signal restricted to section edges, necrotic regions or a positive no-primary control as suspect, and investigate endogenous peroxidase if DAB appears without primary antibody (standard IHC practice). Cytoplasmic staining requires context because HAT1 also has cytoplasmic and mitochondrial annotations; a strong isolated membrane rim is inconsistent with its lack of a transmembrane segment (UniProt O14929).
Boster reagents
Best HAT1 / Histone acetyltransferase type B catalytic subunit IHC Antibodies
Catalog anti-HAT1 antibodies have IHC images from human liver cancer, human lung cancer, mouse bladder, and rat thymus paraffin sections; IF imaging is shown in HeLa cells (catalog image captions).
Real IHC data
Anti-KAT1 / HAT1 Rabbit Monoclonal Antibody
Cat # M03596
Real IHC data
Anti-KAT1/HAT1 Antibody ®
Cat # A03596-2
M03596 has IHC images from human liver cancer, mouse bladder, and rat thymus paraffin sections (M03596 IHC captions). A03596-2 has IHC images from human liver and lung cancer paraffin sections and an IF image from HeLa cells (A03596-2 IHC and IF captions).
Which to pick: For tissue IHC, choose M03596 when a rabbit monoclonal antibody and illustrated staining across human, mouse, and rat paraffin sections are useful; its pictured protocol used EDTA retrieval at pH 8.0 and 1:50 primary antibody (catalog entry; M03596 IHC captions). For IF/ICC, choose A03596-2 because its IF image documents HeLa cells stained at 5 μg/mL (A03596-2 IF caption). For cross-species tissue IHC, M03596 has the broader illustrated evidence; both catalog entries list human, mouse, and rat reactivity, but the fixative is unreported in their paraffin-section IHC captions (catalog entries; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14929 (HAT1_HUMAN, Histone acetyltransferase type B catalytic subunit).
- Human Protein Atlas. HAT1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HAT1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. HAT1 antibody validation summary (2 antibodies).
- Histopathological and epigenetic changes in myocardium associated with cancer therapy-related cardiac dysfunction. ESC heart failure 2022 — PMC9715834.
- Decreased HAT1 expression in granulosa cells disturbs oocyte meiosis during mouse ovarian aging. Reproductive biology and endocrinology : RB&E 2023 — PMC10617186.
- Overexpressed histone acetyltransferase 1 regulates cancer immunity by increasing programmed death-ligand 1 expression in pancreatic cancer. Journal of experimental & clinical cancer research : CR 2019 — PMC6359760.
- Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle. Oncology letters 2025 — PMC12490291.
- PubMed PMID:9427644 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HAT1 (Histone acetyltransferase type B catalytic subunit). Expected localisation Predominantly nuclear in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03596). Validated IHC antibody: M03596 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet M03596); 2. Fixation: Image fixative and duration unreported (datasheet M03596); 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: EDTA pH 8.0 (datasheet M03596); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03596); 8. Primary antibody: Rabbit monoclonal (clone 21H24) anti-HAT1, 1:50 (datasheet M03596); 9. Primary incubation: Overnight at 4 °C (datasheet M03596). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms (A/B); epitope impact unknown (UniProt); Regulation: Oxidative stress enhances nuclear localisation (UniProt). Evidence: UniProt O14929, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet M03596, PMC9715834, PMC10617186. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HAT1/immunohistochemistry-ihc-guide.