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HDAC1 / Histone deacetylase 1 · IHC design guide
Design Immunohistochemistry for HDAC1
Plan chromogenic HDAC1 IHC in paraffin sections with the IHC-validated antibody at 2 μg/ml (datasheet A00256-4). Assess nuclear staining and compare high-staining bone marrow hematopoietic cells with skeletal myocytes, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HDAC1 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 | Nuclear (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00256-4) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent between sections (standard IHC practice; not target-specific) | |
| Caveat | Glia and skeletal myocytes may lack detectable staining (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | No annotated isoforms; one 1–482 chain (UniProt) |
Section 1
Recommended HDAC1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A00256-4) is accompanied by three published HDAC1 IHC procedures (PMC6113427; PMC7471029; PMC3092484).
| Sample | Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A00256-4) |
| Fixation | Image fixative and duration unreported (datasheet A00256-4); 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 A00256-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00256-4) |
| Primary antibody | Rabbit anti-HDAC1, 2 μg/ml (datasheet A00256-4) |
| Primary incubation | Overnight at 4 °C (datasheet A00256-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A00256-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HDAC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval in EDTA pH 8.0 (datasheet: A00256-4). For optimization, compare citrate pH 6.0 at 121°C for 2 min 30 s (PMC7471029: retrieval).
Section 2
What Is the Expected HDAC1 Staining Pattern?
HDAC1 should stain nuclei across many cell types (UniProt Q13547: nucleus; HPA: ubiquitous nuclear expression). Strong examples include colon glandular cells and bone marrow hematopoietic cells (HPA: High). Read the pattern cell by cell: HPA also reports cell populations with low or undetected staining. The tissue IHC profile has Enhanced reliability, reflecting high consistency between antibody staining and RNA expression (HPA: Enhanced). HDAC1 has no transmembrane segment (UniProt Q13547: topology).
| Clear nuclear chromogen in colon glandular cells or bone marrow hematopoietic cells, with little signal outside nuclei. | This matches the expected compartment and two High staining examples (UniProt Q13547: nucleus; HPA: High in both cell populations). Judge intensity in identified cells, rather than averaging across the whole section. |
| Cytoplasmic or membrane staining dominates while nuclei remain pale. | This conflicts with the annotated nuclear location and HPA nuclear tissue profile (UniProt Q13547: nucleus; HPA: ubiquitous nuclear expression). Treat it as suspect staining; review controls and detection before assigning HDAC1 positivity. |
| Strong signal appears in skeletal myocytes, while an expected positive cell population is unstained. | HPA reports HDAC1 as Not detected in skeletal myocytes (HPA: skeletal muscle). This combination raises concern for cross-reactivity or endogenous detection activity, but one unexpected cell population alone does not establish the cause. |
| A uniform brown haze covers nuclei, cytoplasm, and empty areas of the section. | A haze that does not follow cell boundaries or nuclei is difficult to score as HDAC1 (UniProt Q13547: nucleus; standard IHC practice). Check the negative detection control and background before interpreting weak nuclear color. |
| No nuclear signal appears in colon glandular cells or bone marrow hematopoietic cells. | These are High staining populations in HPA tissue IHC (HPA: colon glandular cells; bone marrow hematopoietic cells). A blank result warrants a workflow check; it does not by itself show that the specimen lacks HDAC1. |
💡Expected HDAC1 appearanceCall a convincing positive result when identifiable cells show predominantly nuclear staining, strong in HPA High populations such as colon glandular cells; dominant cytoplasmic color or a uniform section-wide haze is suspect (UniProt Q13547: nucleus; HPA: colon glandular cells, High; standard IHC practice).
| Cell-specific tissue pattern | The profile is broadly nuclear, yet levels vary by cell type (HPA: ubiquitous nuclear expression). HPA reports cardiomyocytes Low and skeletal myocytes Not detected; do not score an entire tissue from its name alone (HPA: heart muscle; skeletal muscle). |
| IHC antibody evidence | HPA029693, CAB005017, and CAB068191 each have Enhanced IHC validation (HPA: antibody validation). This supports their reported staining patterns; it does not guarantee that every chromogenic signal in a new specimen is specific. |
| Topology and processing | HDAC1 is annotated as nuclear, without a transmembrane segment, signal peptide, or propeptide; its annotated chain spans residues 1–482 (UniProt Q13547). These annotations give no basis to expect a membrane or shed-protein staining pattern. |
| Epitope and fixation limits | UniProt lists modified residues, but the supplied evidence gives no antibody epitope or target-specific fixation sensitivity (UniProt Q13547: modified residues; HPA: antibody validation). Do not predict retrieval performance from those annotations. |
| IF/ICC Q&A: where should signal appear? | Mainly in the nucleoplasm (HPA: subcellular, Nucleoplasm enhanced). This is a localisation cross-check for the separate IF/ICC guide; HPA tissue IHC supplies the paraffin-section cell-pattern evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive area is blank. | The staining or detection workflow may have failed; colon glandular cells are High in HPA tissue IHC (HPA: colon). | Check section integrity, antigen retrieval, primary-antibody application, detection reagents, and counterstain on a control section (standard IHC practice). Reassess the same identified cells. |
| Nuclei are pale but cytoplasm is dark. | The dominant compartment conflicts with HDAC1 localisation (UniProt Q13547: nucleus; HPA: ubiquitous nuclear expression). | Compare with a negative detection control, inspect chromogen deposition, and repeat with an IHC-validated antibody if needed (HPA: IHC Enhanced; standard IHC practice). |
| Brown color persists without primary antibody. | Endogenous detection activity or nonspecific detection-reagent binding may contribute (standard IHC practice). | Use the appropriate endogenous-activity block and review detection controls before scoring weak signal (standard IHC practice). HPA staining levels do not identify the source of this background. |
| Color is diffuse across cells and blank spaces. | Nonlocalized background obscures a nuclear readout (UniProt Q13547: nucleus; standard IHC practice). | Review blocking, washes, and detection conditions with matched controls; score HDAC1 only where cell-associated nuclear signal remains distinguishable (standard IHC practice). |
| A presumed negative tissue contains a few positive nuclei. | HPA Not detected calls apply to named cell populations, such as skeletal myocytes or cerebral-cortex glia, not every cell in those tissues (HPA: skeletal muscle; cerebral cortex). | Identify the stained cell type and compare its nuclear pattern with an HPA High population before calling the result discordant (HPA: colon glandular cells, High; standard IHC practice). |
| Heart muscle looks weaker than expected from the UniProt tissue description. | UniProt reports higher levels in heart, while HPA tissue IHC calls cardiomyocytes Low (UniProt Q13547: tissue specificity; HPA: heart muscle). The statements describe different evidence and levels of resolution. | Use the HPA cardiomyocyte observation for this cell-level IHC comparison; evaluate nuclear staining and controls before changing the assay (HPA: cardiomyocytes, Low; UniProt Q13547: nucleus). |
Experiment evidence
IHC & IF images showing HDAC1 staining
A00256-4 · IHCIHC analysis of HDAC1 using anti-HDAC1 antibody (A00256-4). HDAC1 was detected in a paraffin-embedded section of human esophageal squamous c
A00256-4 · IHCIHC analysis of HDAC1 using anti-HDAC1 antibody (A00256-4). HDAC1 was detected in a paraffin-embedded section of human lung cancer tissue. H
M00256-2 · IHCHuman tonsil was stained with anti-HDAC1 rabbit antibody
M00256-2 · IHCMouse liver was stained with anti-HDAC1 rabbit antibody
M00256 · IHCImmunohistochemical analysis of paraffin-embedded human testis, using HDAC1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000116478-HDAC1/tissue/cerebral+cortex#imid_18803482
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000116478-HDAC1/tissue/colon#imid_18799552
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000116478-HDAC1/tissue/liver#imid_18799553
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000116478-HDAC1/tissue/kidney#imid_18799554
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029693; green = HDAC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029693; green = HDAC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HDAC1 IHC & IF
🧪Run bone marrow first: hematopoietic cells should show nuclear HDAC1 staining (HPA: High in bone marrow hematopoietic cells; UniProt Q13547: Nucleus). Use skeletal muscle myocytes as a low-signal comparison (HPA: Not detected in skeletal muscle myocytes); mature erythrocytes within the marrow section lack nuclei and should show no nuclear signal (standard histology).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HDAC1 in A-431, U-251MG, U2OS, U2OS, siRNA 2 (10x), U2OS, scrambled (10x), U2OS, siRNA 1 (10x), HeLa BAC 6203, HeLa , with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; rabbit IgG matched to the primary antibody’s isotype and clonality where known; and HDAC1-knockout material as a biological negative (caption: rabbit anti-HDAC1; standard IHC controls). Quench endogenous peroxidase and inspect marrow for background from blood cells before scoring nuclear DAB staining (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A00256-4 paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU caption: heat retrieval in EDTA; HPA: ICC-IF images available). In bone marrow, endogenous peroxidase can complicate chromogenic interpretation, so assess the no-primary slide alongside nuclear staining (standard IHC practice).
HPA tissue IHC evidence for HDAC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 | Respiratory epithelial cells | 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HDAC1 IHC Tips
Troubleshoot HDAC1 staining in paraffin sections by checking retrieval, nuclear localisation, assay controls and scoring before interpreting differences between samples.
What retrieval should I start with if HDAC1 nuclear staining is weak?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet A00256-4). The pictured section of human esophageal squamous carcinoma was stained after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (caption A00256-4). If nuclear signal is weak, check that heating reaches the sections uniformly and compare retrieval duration on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Judge improvement by crisp nuclear staining with limited extranuclear deposit, because HDAC1 is nuclear and mainly nucleoplasmic (UniProt Q13547; HPA subcellular).
Could fixation explain poor HDAC1 staining in my paraffin sections?
Target-specific sensitivity to fixation is unknown: the selected tissue caption says only that the section was paraffin embedded and does not state its fixative (caption A00256-4). Record the fixative, fixation interval and processing history for each specimen, then compare sections processed alike before changing retrieval or antibody concentration (standard IHC practice). Excessive or uneven fixation can alter antigen accessibility in paraffin IHC generally, but no HDAC1-specific fixation threshold follows from the supplied evidence (standard IHC practice). Use nuclear morphology and a matched positive control to distinguish a processing problem from weak expression; the expected HDAC1 compartment is the nucleus (UniProt Q13547).
How should I troubleshoot HDAC1 staining outside nuclei?
Expect predominantly nuclear, especially nucleoplasmic, staining because HDAC1 is annotated in the nucleus and the cellular imaging summary identifies nucleoplasm (UniProt Q13547; HPA subcellular). If brown signal spreads through cytoplasm or along section edges, inspect the counterstain and tissue morphology, then repeat with a no-primary control to assess detection background (standard IHC practice). HDAC1 has no annotated transmembrane segment, so a continuous membrane rim is inconsistent with its supplied localisation evidence (UniProt Q13547 topology; HPA subcellular). Compare well-preserved internal nuclei with an appropriate reference section before calling diffuse extranuclear staining a biological redistribution (standard IHC practice).
Could an isoform or masked epitope explain inconsistent HDAC1 IHC?
The supplied HDAC1 record lists 0 annotated isoforms and one chain spanning residues 1–482; it does not identify this antibody’s binding epitope (UniProt Q13547; caption A00256-4). Therefore, do not assign patchy nuclear staining to a particular isoform or modification without independent epitope and specificity evidence (UniProt Q13547; standard IHC practice). Annotated modifications include acetylation, S-nitrosylation and phosphorylation, but their effects on this antibody’s tissue staining are unreported (UniProt Q13547). When staining varies, first compare matched processing and EDTA pH 8.0 retrieval, then assess antibody specificity with an appropriate independent control (datasheet A00256-4; standard IHC practice).
How can I check HDAC1 localisation by IF alongside this IHC assay?
Use IF as an independent localisation check: HDAC1 is mainly nucleoplasmic, and the supplied cellular imaging record includes ICC/IF images (HPA subcellular). Multiplex with a marker identifying the cell population being assessed, such as a validated epithelial marker when examining the documented esophageal squamous cells, and inspect overlap within counterstained nuclei (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low background in the specimen, checking tissue autofluorescence and single-label controls before interpreting overlap (standard IF practice). Because HDAC1 is nuclear and has no transmembrane segment, permeabilisation must permit antibody access to a nuclear epitope; optimise it for the fixation used (UniProt Q13547; standard IF practice).
What should I check when HDAC1 DAB staining looks diffuse?
The selected paraffin-section image used 10% goat serum blocking, a peroxidase-linked secondary antibody and DAB development (caption A00256-4). For diffuse brown deposit, inspect a no-primary control, confirm that the peroxidase block and washes are effective, and shorten chromogen development if background obscures nuclei (standard IHC practice). Compare adjacent intact cells: convincing HDAC1 signal should resolve within nuclei rather than form a uniform coating over tissue or an edge-only deposit (UniProt Q13547; HPA subcellular; standard IHC practice). Keep the reported 2 μg/ml primary condition as a reference while varying one background-control step at a time (caption A00256-4; standard IHC practice).
How should I quantify HDAC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score HDAC1 in the nuclei of a defined, viable cell population, since its reported tissue and cellular staining is nuclear or nucleoplasmic (HPA tissue IHC; HPA subcellular). Record the percentage of positive nuclei and intensity, or calculate a nuclear H-score from intensity categories for a 0–300 scale (standard IHC practice). Normalise counts to the number of evaluable target-cell nuclei, or report positive nuclei per mm² of the same defined compartment, excluding necrosis and section edges (standard IHC practice). Apply identical retrieval, detection, counterstain and scoring thresholds across samples, and report cell type because HPA levels vary between annotated cell populations (datasheet A00256-4; HPA tissue IHC; standard IHC practice).
How can I distinguish true HDAC1 positivity from staining artefact?
A credible positive has interpretable nuclear staining in preserved cells, consistent with HDAC1’s nuclear localisation and the reported ubiquitous nuclear tissue profile (UniProt Q13547; HPA tissue IHC). Check the cell identity: HPA reports high staining in esophageal squamous epithelial cells but no detection in skeletal-muscle myocytes, so pooling unlike cells can mislead (HPA tissue IHC). Treat edge-heavy deposit, staining over necrosis, or predominantly membrane or cytoplasmic colour as suspect, and compare a no-primary control for endogenous enzyme or detection artefact (UniProt Q13547 topology; standard IHC practice). Interpret intensity only after confirming matched processing and the documented EDTA pH 8.0 retrieval (datasheet A00256-4; standard IHC practice).
Boster reagents
Best HDAC1 / Histone deacetylase 1 IHC Antibodies
The catalog lists three IHC antibodies with human tissue images (catalog IHC captions); two also list IF, and catalog reactivity spans human, mouse and rat (catalog applications/reactivity).
Real IHC data
Anti-HDAC1 Antibody ®
Cat # A00256-4
Real IHC data
Anti-HDAC1 Rabbit Monoclonal Antibody
Cat # M00256-2
Real IHC data
Anti-HDAC1 Rabbit Monoclonal Antibody
Cat # M00256
A00256-4 has an IHC image of a human esophageal squamous carcinoma paraffin section, and M00256 has one of a human testis paraffin section (respective IHC captions). M00256-2 has IHC images of human tonsil and mouse and rat liver (M00256-2 IHC captions).
Which to pick: For human paraffin-section IHC, choose A00256-4 if its documented EDTA retrieval at pH 8.0 and 2 μg/ml overnight incubation suit the assay (A00256-4 IHC caption); M00256 is another option shown on a human testis paraffin section (M00256 IHC caption). For IF/ICC, choose rabbit monoclonal M00256, which lists both applications (M00256 catalog applications/clone); for cross-species IHC or IF, choose rabbit monoclonal M00256-2, which lists human, mouse and rat reactivity and has IHC images from all three species (M00256-2 catalog applications/reactivity; IHC captions). Fixative is unreported for all three IHC examples, and the M00256-2 captions do not specify tissue processing (respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13547 (HDAC1_HUMAN, Histone deacetylase 1).
- Human Protein Atlas. HDAC1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HDAC1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. HDAC1 antibody validation summary (3 antibodies).
- Role of histone deacetylase 1 in distant metastasis of pancreatic ductal cancer. Cancer science 2018 — PMC6113427.
- Aberrant HDAC3 expression correlates with brain metastasis in breast cancer patients. Thoracic cancer 2020 — PMC7471029.
- Suppression of histone deacetylase 1 by JSL-1 attenuates the progression and metastasis of cholangiocarcinoma via the TPX2/Snail axis. Cell death & disease 2022 — PMC8993895.
- Hyperoxia impairs alveolar formation and induces senescence through decreased histone deacetylase activity and up-regulation of p21 in neonatal mouse lung. Pediatric research 2011 — PMC3092484.
- PubMed PMID:8602529 — UniProt-cited evidence.
- PubMed PMID:8646880 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HDAC1 (Histone deacetylase 1). Expected localisation Nuclear (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00256-4). Validated IHC antibody: A00256-4 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A00256-4); 2. Fixation: Image fixative and duration unreported (datasheet A00256-4); 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 A00256-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00256-4); 8. Primary antibody: Rabbit anti-HDAC1, 2 μg/ml (datasheet A00256-4); 9. Primary incubation: Overnight at 4 °C (datasheet A00256-4). Watch for: Fixation: Keep fixation consistent between sections (standard IHC practice; not target-specific); Caveat: Glia and skeletal myocytes may lack detectable staining (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–482 chain (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q13547, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A00256-4, PMC6113427, PMC7471029. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HDAC1/immunohistochemistry-ihc-guide.