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ADH1A / Alcohol dehydrogenase 1A · IHC design guide
Design Immunohistochemistry for ADH1A
Plan ADH1A staining in paraffin sections around selective hepatocyte cytoplasmic expression (HPA tissue IHC). This guide covers IHC handling and interpretation, including the possibility that staining includes protein from other genes (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ADH1A 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 | Hepatocyte cytoplasm (HPA tissue IHC) | |
| Staining pattern | Selective hepatocyte cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB10029) | |
| Positive control | Appendix+3 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 may include protein from other genes (HPA tissue IHC) | |
| Regulation | Regulation not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans aa 2–375 (UniProt) |
Section 1
Recommended ADH1A IHC & IF Protocols
The catalog antibody protocol uses EDTA retrieval (datasheet PB10029). The published IHC protocols below provide paraffin-section conditions for human and murine tissues (PMC13357096; PMC13326605).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet PB10029) |
| Fixation | Image fixative and duration unreported (datasheet PB10029); 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 PB10029); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB10029) |
| Primary antibody | Rabbit anti-ADH1A, 2-5μg/ml (datasheet PB10029) |
| Primary incubation | Overnight at 4 °C (datasheet PB10029) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB10029) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ADH1A-positive staining in endocrine cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Selective cytoplasmic expression in hepatocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet PB10029); citrate pH 6.0 is a published alternative (PMC13357096).
Section 2
What Is the Expected ADH1A Staining Pattern?
ADH1A is a cytoplasmic protein with no transmembrane segment (UniProt P07327). In paraffin-section IHC, expect selective cytoplasmic staining of hepatocytes, reported at medium intensity; the HPA tissue profile has Enhanced reliability (HPA tissue IHC). Medium staining is also reported in appendix and duodenal endocrine cells and small-intestinal enterocytes (HPA tissue IHC). Interpret these patterns cautiously because the HPA tissue assessment includes antibodies targeting proteins from more than one gene (HPA tissue IHC).
| Hepatocytes show medium cytoplasmic chromogen, with cell boundaries and nuclei distinguishable. | This matches the principal tissue pattern (HPA tissue IHC: medium in hepatocytes) and cytoplasmic location (UniProt P07327). Score cytoplasmic intensity and the fraction of stained hepatocytes separately (general IHC practice). |
| Signal is predominantly nuclear or sharply outlines hepatocyte membranes, without convincing cytoplasmic staining. | That distribution conflicts with the expected cytoplasmic IHC pattern (HPA tissue IHC; UniProt P07327). Check the counterstain, detection controls and antibody performance before assigning ADH1A localisation (general IHC practice). |
| Strong signal appears in a cell population reported as undetected, such as adipocytes. | Adipocytes are reported as not detected (HPA tissue IHC). Consider nonspecific antibody binding, recognition of a related protein, or endogenous chromogenic activity; the HPA antibody assessment cautions that more than one gene may be targeted (HPA tissue IHC). |
| Chromogen is diffuse across tissue or appears in the negative control. | This does not establish cell-specific ADH1A staining (general IHC practice). Review primary-antibody omission controls, blocking, washes and detection activity, then reassess whether hepatocyte cytoplasm stands above background (general IHC practice; HPA tissue IHC). |
| A liver section shows no hepatocyte signal. | Liver hepatocytes are a reported positive population at medium intensity (HPA tissue IHC). First check tissue preservation, assay controls and detection steps; absence of staining in one section alone cannot establish absent ADH1A expression (general IHC practice). |
💡Expected ADH1A appearanceCall positive when hepatocyte cytoplasm has discernible, approximately medium staining (HPA tissue IHC; UniProt P07327); isolated nuclear, membrane-only or control-matched diffuse colour warrants investigation (general IHC practice).
| Cell and tissue context | Liver is RNA tissue-enriched, while hepatocytes show medium protein staining (HPA tissue IHC). Appendix and duodenal endocrine cells and small-intestinal enterocytes are additional medium-staining populations (HPA tissue IHC); identify the cell type before comparing slides (general IHC practice). |
| Antibody specificity and validation | The tissue profile has Enhanced reliability but cautions that antibodies target proteins from more than one gene (HPA tissue IHC). CAB009562 has Enhanced IHC validation; HPA047814 and HPA060902 are Supported (HPA antibodies). These grades do not establish gene-exclusive staining (HPA tissue IHC). |
| Subcellular expectation | A cytoplasmic IHC pattern accords with UniProt localisation and lack of a transmembrane segment (UniProt P07327). The HPA ICC-IF summary supports cytosol but labels plasma membrane and several additional sites uncertain, with a multiple-gene antibody caution (HPA subcellular ICC-IF). |
| IF/ICC: what localisation should be expected? | Cytosol is supported in the HPA ICC-IF assessment; plasma membrane is uncertain, as are nucleoplasm and ciliary locations (HPA subcellular ICC-IF). Treat those additional signals as provisional, particularly because the antibodies may target proteins from multiple genes (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in liver hepatocytes | The expected medium hepatocyte signal is missing (HPA tissue IHC); the assay may have failed (general IHC practice). | Check positive and negative controls, reagent order, antigen-retrieval conditions and detection chemistry before interpreting the specimen (general IHC practice). |
| Signal is widespread across unrelated cell types | The distribution exceeds the selective hepatocyte profile (HPA tissue IHC); nonspecific binding or recognition of another gene product is possible (general IHC practice; HPA tissue IHC caution). | Compare cell-resolved staining with an appropriate negative control and, if available, an independently validated antibody (general IHC practice). |
| Diffuse chromogen persists without primary antibody | Primary-independent colour suggests endogenous detection activity or detection-reagent background (general IHC practice). | For an HRP chromogen, check the endogenous-peroxidase block; review detection-only controls, blocking and washes (general IHC practice). |
| Nuclear or membrane-only signal dominates | This conflicts with cytoplasmic localisation (UniProt P07327; HPA tissue IHC); membrane localisation in ICC-IF is uncertain (HPA subcellular ICC-IF). | Recheck compartment assignment with the counterstain and controls; avoid scoring that pattern as confirmed ADH1A (general IHC practice). |
| Only faint cytoplasmic colour is present | The reported hepatocyte level is medium, but observed intensity depends on assay conditions (HPA tissue IHC; general IHC practice). | Compare a known-positive liver control in the same run; review antibody dilution, retrieval and detection settings without assigning a target-specific optimum (general IHC practice). |
| An unexpected cell population stains alongside hepatocytes | The HPA profile includes other medium-positive populations and warns of multiple-gene antibody targeting (HPA tissue IHC). | Identify the stained cell type first, compare it with the HPA tissue pattern and controls, and report unresolved staining as uncertain (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ADH1A staining
PB10029 · IHCIHC analysis of ADH1A using anti-ADH1A antibody (PB10029). ADH1A was detected in a paraffin-embedded section of human liver cancer tissue. H
PB10029 · IHCIHC analysis of ADH1A using anti-ADH1A antibody (PB10029). ADH1A was detected in a paraffin-embedded section of mouse liver tissue. Heat med
PB10029 · IFIF analysis of ADH1A using anti-ADH1A antibody (PB10029). ADH1A was detected in immunocytochemical section of U20S cells. Enzyme antigen ret
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000187758-ADH1A/tissue/cerebral+cortex#imid_17502999
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000187758-ADH1A/tissue/colon#imid_17502925
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000187758-ADH1A/tissue/liver#imid_17502910
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000187758-ADH1A/tissue/pancreas#imid_17502869
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA047814; green = ADH1A, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA047814; green = ADH1A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ADH1A IHC & IF
🧪Run liver first: hepatocytes should stain at the Medium level (HPA: Liver, hepatocytes, Medium). Use adipose tissue as the negative comparator (HPA: Adipose tissue, adipocytes, Not detected); non-hepatocyte cells on the liver slide provide an internal background comparison, but their ADH1A status is not established by the supplied HPA row.
Positive control tissue: Appendix (Endocrine cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADH1A in ASC52telo, SiHa, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Plasma membrane (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-, isotype-, and clonality-matched rabbit IgG controls, plus ADH1A-knockout material as a biological negative (hero caption: rabbit anti-ADH1A antibody). For liver IHC with HRP/DAB detection, block endogenous peroxidase and inspect the controls for residual background (hero caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB10029 tissue-IHC caption does not state the fixative (PB10029 tissue-IHC caption: fixative not stated). The caption provides heat retrieval in EDTA at pH 8.0 as a starting condition, but does not establish that retrieval is required; frozen sections and IF have no demonstrated advantage in the supplied evidence (PB10029 tissue-IHC caption: heat retrieval in EDTA, pH 8.0). In liver, assess cytoplasmic staining against background and endogenous peroxidase signal (UniProt P07327: Cytoplasm; HPA: Liver, hepatocytes, Medium; PB10029 tissue-IHC caption: HRP/DAB detection).
HPA tissue IHC evidence for ADH1A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
| Small intestine | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADH1A IHC Tips
Troubleshoot ADH1A staining in paraffin sections by checking retrieval, cytoplasmic localisation, cell identity, and antibody specificity before interpreting signal intensity.
What retrieval should I use when ADH1A staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet PB10029). The selected paraffin-section example used that retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C (caption PB10029). If staining is weak, keep retrieval conditions consistent while checking section processing, primary incubation, and a positive liver control, rather than changing several variables together (standard IHC practice; HPA: selective cytoplasmic expression in hepatocytes). Compare test sections with a no-primary control to distinguish recovered signal from retrieval-related background (standard IHC practice). Record heating and cooling conditions so a useful result can be reproduced (standard IHC practice).
Could fixation explain weak or uneven ADH1A staining?
The selected ADH1A image shows a paraffin-embedded liver cancer section but does not state its fixative (caption PB10029). Target-specific sensitivity to fixation is therefore unknown from the supplied evidence; neither tissue staining patterns nor protein features establish it (caption PB10029; HPA: tissue IHC profile; UniProt P07327). For uneven staining, compare regions with intact morphology, review the specimen’s recorded fixation and processing history, and check whether the pattern tracks section edges (standard IHC practice). Run a similarly processed positive control alongside the test section, keeping retrieval and detection conditions matched (standard IHC practice). Report the actual fixative when known rather than treating paraffin embedding as proof of formalin fixation (standard IHC practice).
Where should ADH1A staining appear, and how should membrane signal be judged?
Expect predominantly cytoplasmic staining in hepatocytes when assessing liver sections (UniProt P07327: cytoplasm; HPA: selective cytoplasmic expression in hepatocytes). ADH1A has no annotated transmembrane segment, so crisp membrane-only staining warrants additional specificity checks (UniProt P07327 topology). HPA reports supported cytosolic localisation, but labels its plasma-membrane assignment uncertain and cautions that the underlying antibodies can target proteins from multiple genes (HPA: subcellular). Examine the DAB signal against cell boundaries and the counterstain, then compare it with a no-primary control (standard IHC practice). Do not assign uncertain nucleoplasmic or ciliary signals to ADH1A from chromogenic appearance alone (HPA: subcellular; standard IHC practice).
Can this staining distinguish ADH1A from related alcohol dehydrogenases?
Do not infer ADH1A-specific detection from staining pattern alone: ADH1A can form dimers with closely related ADH1B and ADH1C subunits (UniProt P07327 subunit). UniProt lists 0 alternative ADH1A isoforms, but that does not establish which related proteins an antibody recognises (UniProt P07327 isoforms; standard IHC practice). Review the catalog antibody’s documented epitope and cross-reactivity, if available, and use independent target-specific validation before assigning a signal uniquely to ADH1A (standard IHC practice). This matters because HPA’s enhanced tissue-staining assessment explicitly cautions that its antibody targets proteins from more than one gene (HPA: tissue IHC). Report unresolved staining as antibody-dependent signal rather than isoform-specific expression (standard IHC practice).
How should I check an ADH1A IHC pattern with immunofluorescence?
Use IF as a complementary localisation check, while keeping the paraffin-section chromogenic result as the primary readout (standard IHC/IF practice). In liver, multiplex ADH1A with a validated hepatocyte marker and inspect whether cytosolic signal falls within marker-positive cells (HPA: selective cytoplasmic expression in hepatocytes; UniProt P07327: cytoplasm; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence in an unstained control, and separate channels to limit bleed-through (standard IF practice). Permeabilise sufficiently for an intracellular epitope, then compare permeabilised and control preparations because ADH1A is cytoplasmic and lacks a transmembrane segment (UniProt P07327 topology; standard IF practice). Do not transfer the paraffin IHC incubation settings to IF without validation (caption PB10029; standard IF practice).
How can I separate ADH1A DAB signal from background?
Inspect no-primary and secondary-only controls alongside the stained section to locate nonspecific detection signal (standard IHC practice). In a peroxidase-DAB workflow, use an appropriate endogenous peroxidase block and assess whether pigment or residual enzyme activity persists in controls (caption PB10029: peroxidase detection and DAB; standard IHC practice). The selected image used 10% goat serum blocking and 2 μg/ml primary antibody, providing documented starting conditions for that antibody (caption PB10029). If diffuse staining rises as primary concentration increases, titrate the antibody and compare cytoplasmic signal with background in the same run (standard IHC practice; UniProt P07327: cytoplasm). Check section edges and damaged areas before calling dark DAB deposits positive (standard IHC practice).
What is a defensible way to score ADH1A IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; liver assessment should focus on hepatocyte cytoplasm (HPA: selective cytoplasmic expression in hepatocytes; UniProt P07327: cytoplasm). Report the percentage of positive eligible cells and an intensity-based H-score from 0–300, with fixed thresholds across the batch (standard IHC practice). Normalise counts to the number of viable eligible cells; if measuring stained area or cell density, state the analysed viable tissue area in mm² (standard IHC practice). Exclude folds, edges, necrosis, and pigment identified by controls, and use matched acquisition or scoring settings (standard IHC practice). Record whether results describe tumour cells, adjacent hepatocytes, or both when those compartments are present (standard IHC practice).
When is an apparent ADH1A-positive region likely to be artefact?
Give greatest weight to cell-associated cytoplasmic staining in morphologically intact hepatocytes (HPA: selective cytoplasmic expression in hepatocytes; UniProt P07327: cytoplasm; standard IHC practice). Treat membrane-only or strongly nuclear patterns cautiously because UniProt places ADH1A in the cytoplasm, while HPA marks membrane and nucleoplasmic assignments uncertain (UniProt P07327; HPA: subcellular). Staining limited to section edges, necrotic regions, or cells outside the expected population needs comparison with morphology and controls (HPA: tissue IHC; standard IHC practice). DAB deposited in a no-primary control can indicate endogenous peroxidase activity or other detection background (caption PB10029: peroxidase-DAB workflow; standard IHC practice). Even convincing staining cannot by itself resolve ADH1A from related-gene reactivity (HPA: tissue IHC caution).
Boster reagents
Best ADH1A / Alcohol dehydrogenase 1A IHC Antibodies
PB10029 has IHC images from paraffin sections of human liver cancer and mouse and rat liver, plus an IF image from U20S cells (catalog image captions).
Real IHC data
Anti-Alcohol Dehydrogenase/ADH1A Antibody ®
Cat # PB10029
PB10029 is the card that will render; its IHC captions show staining in paraffin sections of human liver cancer and mouse and rat liver (catalog IHC captions). Its IF caption shows staining in U20S cells (catalog IF caption).
Which to pick: Choose PB10029 for tissue IHC because its own images show paraffin sections stained at 2 μg/ml after EDTA retrieval (PB10029 IHC captions). Choose PB10029 for IF/ICC and for work across human, mouse and rat because those applications and species are listed for it; the supplied IF image is from U20S cells (PB10029 catalog applications, reactivity and IF caption). The IHC captions do not report the fixative (PB10029 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07327 (ADH1A_HUMAN, Alcohol dehydrogenase 1A).
- Human Protein Atlas. ADH1A tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ADH1A subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the nucleoplasm, primary cilium, primary cilium transition zone and basal body. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. ADH1A antibody validation summary (3 antibodies).
- Overexpression of alcohol dehydrogenase 1 A inhibits the progress of triple negative breast cancer via Wnt/β-catenin signaling. Scientific reports 2025 — PMC12475006.
- A pan-cancer functional atlas of ADH1A reveals its conserved role in the tumor microenvironment and immunity. Translational cancer research 2026 — PMC13357096.
- mTOR/HDAC1 Crosstalk Mediated Suppression of ADH1A and ALDH2 Links Alcohol Metabolism to Hepatocellular Carcinoma Onset and Progression in silico. Frontiers in oncology 2019 — PMC6787164.
- HBx-mediated GPT2 suppression promotes liver cancer development by downregulating ADH1A. Cancer biology & medicine 2026 — PMC13326605.
- PubMed PMID:3013304 — UniProt-cited evidence.
- PubMed PMID:2935875 — UniProt-cited evidence.
- PubMed PMID:2920825 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ADH1A (Alcohol dehydrogenase 1A). Expected localisation Hepatocyte cytoplasm (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB10029). Validated IHC antibody: PB10029 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine 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 PB10029); 2. Fixation: Image fixative and duration unreported (datasheet PB10029); 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 PB10029); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB10029); 8. Primary antibody: Rabbit anti-ADH1A, 2-5μg/ml (datasheet PB10029); 9. Primary incubation: Overnight at 4 °C (datasheet PB10029). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining may include protein from other genes (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans aa 2–375 (UniProt); Regulation: Regulation not annotated (UniProt). Evidence: UniProt P07327, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet PB10029, PMC13357096, PMC13326605. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ADH1A/immunohistochemistry-ihc-guide.