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ADH1C / Alcohol dehydrogenase 1C · IHC design guide
Design Immunohistochemistry for ADH1C
Plan chromogenic ADH1C IHC using cytoplasmic staining in gastrointestinal glandular cells and strong hepatocyte staining as reference patterns (HPA tissue IHC). The catalog antibody has an IHC dilution range of 1:50–1:200 (datasheet); interpret staining with the HPA warning that antibodies may detect proteins from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ADH1C Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal mainly in GI glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Liver+4 more · see all | |
| Negative control | Bronchus+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; mature chain spans residues 2–375 (UniProt) |
Section 1
Recommended ADH1C IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with four published ADH1C IHC protocols (PMC13084684; PMC9855755; PMC8921497; PMC7438573).
| Sample | Paraffin-embedded human esophagus tissue; fixative not specified (datasheet A02650) |
| Fixation | Image fixative and duration unreported (datasheet A02650); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-ADH1C, 1:50-1:200 (datasheet A02650) |
| 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 | ADH1C-positive staining in hepatocytes of liver (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in glandular cells of gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); adjust to the selected article’s conditions when reproducing its protocol.
Section 2
What Is the Expected ADH1C Staining Pattern?
ADH1C is a cytoplasmic protein without a transmembrane segment (UniProt P00326: subcellular location and topology). In paraffin section IHC, expect cytoplasmic staining, especially in hepatocytes, which show High staining; the HPA profile also describes expression mainly in gastrointestinal glandular cells (HPA: tissue IHC). Interpret the pattern cautiously: HPA rates tissue staining Supported, reports medium agreement with RNA data, and warns that the antibodies target proteins from more than one gene (HPA: tissue IHC reliability).
| Hepatocytes show strong cytoplasmic staining, while surrounding structures remain distinguishable on the counterstained section. | This matches the reported High hepatocyte signal and cytoplasmic location (HPA: liver tissue IHC; UniProt P00326: subcellular location). Assess cell identity and compartment together; a dark area without recognizable stained hepatocytes is less persuasive than a clear cellular pattern (general IHC practice). |
| Signal appears predominantly nuclear, or outlines cell borders without appreciable cytoplasmic staining. | This does not match the expected tissue IHC pattern (HPA: tissue IHC profile; UniProt P00326: subcellular location). HPA ICC-IF lists plasma membrane as uncertain and nucleoplasm as an additional uncertain location; those observations do not establish a membrane or nuclear IHC pattern (HPA: subcellular ICC-IF). Review staining controls before assigning ADH1C (general IHC practice). |
| Prominent staining occurs in cells scored Not detected by HPA, such as bronchial respiratory epithelium or pancreatic exocrine glandular cells. | Such staining conflicts with those specific HPA tissue observations (HPA: bronchus and pancreas tissue IHC). Possible explanations include nonspecific antibody binding or endogenous chromogenic detection activity (general IHC practice). Because HPA cautions that its antibodies target proteins from more than one gene, staining alone cannot identify which related protein produced the signal (HPA: tissue IHC reliability). |
| Color spreads across extracellular areas, section edges, or many unrelated cell populations, obscuring cellular boundaries. | A diffuse deposit cannot be scored confidently as the reported cytoplasmic cell pattern (HPA: tissue IHC profile). Check the detection and blocking controls, wash conditions, and counterstain; excess background can make a genuinely positive compartment difficult to distinguish (general IHC practice). Do not treat overall section darkness as evidence of ADH1C expression (general IHC practice). |
| Hepatocytes show no detectable signal in a section intended as a positive control. | This disagrees with the reported High hepatocyte staining (HPA: liver tissue IHC). First check section integrity, primary antibody application, detection reagents, and the run controls (general IHC practice). An absent signal in one run does not establish that ADH1C is absent from the sampled hepatocytes; technical performance and tissue quality need assessment (general IHC practice). |
💡Expected ADH1C appearanceCall a result positive when recognizable hepatocytes have clear, predominantly cytoplasmic High staining (HPA: liver tissue IHC; UniProt P00326: subcellular location); widespread haze or isolated nuclear staining is a poor match (general IHC practice; HPA: subcellular ICC-IF, nucleoplasm uncertain).
| Choice of tissue and cell population | Liver hepatocytes provide a reported High staining reference; appendix and duodenum endocrine cells, and gallbladder glandular cells, are reported Medium (HPA: tissue IHC). Compare the same named cell population when judging agreement; different cells within one organ need not have the listed level (general IHC practice). |
| Specificity of the reported tissue pattern | HPA labels tissue IHC Supported and reports medium consistency with RNA expression; it also cautions that the antibodies target proteins from more than one gene (HPA: tissue IHC reliability). Treat a matching pattern as supportive evidence, while recognizing that this record does not prove ADH1C-specific staining (HPA: tissue IHC reliability). |
| Molecular location and related class I enzymes | UniProt places ADH1C in the cytoplasm and reports no transmembrane segment; it can form dimers with class I alcohol dehydrogenase chains ADH1A or ADH1B (UniProt P00326: location, topology, subunit). Dimerization describes the protein complex and does not by itself validate an antibody signal as ADH1C-specific (general IHC interpretation). |
| IF/ICC Q: Should membrane or ciliary staining be expected here? | A: HPA ICC-IF supports cytosol, calls plasma membrane uncertain, and lists primary cilium, transition zone, and basal body as additional uncertain locations; it cautions about antibodies targeting multiple genes (HPA: subcellular ICC-IF). Use those observations only as qualified IF context, not as a paraffin section IHC acceptance criterion (HPA: subcellular ICC-IF; HPA: tissue IHC profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| No hepatocyte staining in the positive reference section | The result conflicts with reported High hepatocyte staining (HPA: liver tissue IHC); a failed staining run or unsuitable section is possible (general IHC practice). | Verify the section, primary antibody step, detection reagents, and run controls; repeat the affected step after identifying a control failure (general IHC practice). Avoid concluding biological absence from this slide alone (general IHC practice). |
| Weak, uneven cytoplasmic signal in liver | Uneven section processing, reagent coverage, or detection can produce patchy staining (general IHC practice); HPA's hepatocyte reference is High (HPA: liver tissue IHC). | Compare intact areas and a concurrently stained control, then review section quality, reagent coverage, and detection performance (general IHC practice). The supplied sources give no ADH1C-specific fixation sensitivity or retrieval setting (UniProt P00326; HPA: tissue IHC). |
| Diffuse brown background obscures cells | Incomplete blocking, inadequate washing, or excess detection signal can reduce contrast in chromogenic IHC (general IHC practice). | Inspect the control without primary antibody, confirm appropriate blocking for the detection system, and optimize washes and detection conditions (general IHC practice). Score only staining that can be assigned to recognizable cells and compartments (general IHC practice). |
| Nuclear or sharply membrane-only staining dominates | This conflicts with cytoplasmic ADH1C and the tissue IHC profile (UniProt P00326: location; HPA: tissue IHC); membrane and nucleoplasm assignments in ICC-IF are uncertain (HPA: subcellular ICC-IF). | Check control sections and detection background, then reassess whether cytoplasmic signal is present (general IHC practice). Do not reinterpret uncertain ICC-IF locations as proof of a nuclear or membrane IHC result (HPA: subcellular ICC-IF). |
| Strong signal appears in a reported negative cell population | For example, HPA reports bronchial respiratory epithelial cells and pancreatic exocrine glandular cells as Not detected (HPA: tissue IHC). Nonspecific binding or endogenous detection activity may contribute (general IHC practice). | Review the control without primary antibody and the relevant blocking steps; compare the exact cell type with HPA's listing (general IHC practice; HPA: tissue IHC). Consider the multiple-gene antibody caution before attributing any residual signal to ADH1C (HPA: tissue IHC reliability). |
| A Medium or Low tissue appears negative while the liver control stains | HPA reports differing levels across cell types, including Medium appendix endocrine cells and Low thyroid glandular cells (HPA: tissue IHC). A weak expected signal may be difficult to separate from background (general IHC practice). | Confirm that the scored cells match the HPA cell population, inspect background and detection performance, and report the observed result with the tissue and cell type (general IHC practice; HPA: tissue IHC). Do not extrapolate the liver intensity to every organ (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing ADH1C staining
A02650 · IHCImmunohistochemistry of paraffin-embedded human esophagus using ADH1C antibody at dilution of 1:100 (40x lens).
A02650 · IHCImmunohistochemistry of paraffin-embedded human stomach using ADH1C antibody at dilution of 1:100 (40x lens).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000248144-ADH1C/tissue/cerebral+cortex#imid_13827745
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000248144-ADH1C/tissue/stomach+1#imid_13836781
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000248144-ADH1C/tissue/colon#imid_13836815
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000248144-ADH1C/tissue/liver#imid_13836816
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA047814; green = ADH1C, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA047814; green = ADH1C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ADH1C IHC & IF
🧪Run liver first: hepatocytes should stain strongly (HPA: High in hepatocytes). Use bronchus respiratory epithelial cells as the negative tissue (HPA: Not detected); on the liver slide, assess nonhepatocyte cells against the no-primary background rather than assuming they are ADH1C-negative (HPA: High in hepatocytes only).
Positive control tissue: Liver (Hepatocytes, HPA High)
Negative control tissue: Bronchus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADH1C in ASC52telo, SiHa, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Plasma membrane (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host species and clonality, and an ADH1C knockout biological negative if available (standard IHC practice). In liver, block endogenous peroxidase for chromogenic detection and distinguish pigment from specific staining (standard IHC practice).
⚠️Feasibility: No matched source reports an ADH1C-specific fixation window or antigen-retrieval dependency; optimize retrieval on paired paraffin sections (supplied target/application evidence). The selected A02650 paraffin-section caption does not report a fixative, so fixation is unconfirmed (selected-SKU tissue-IHC caption). These data do not establish whether frozen sections or IF are easier; expected cytoplasmic staining is consistent with the target annotation (UniProt P00326: Cytoplasm). The selected A02650 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A02650).
HPA tissue IHC evidence for ADH1C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADH1C IHC Tips
Troubleshoot ADH1C staining in paraffin sections by checking cytoplasmic localisation, cell type, antibody specificity and processing controls.
How should I adjust retrieval when ADH1C staining is weak in paraffin sections?
Start with citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). If staining remains weak, compare adjacent sections with a modestly extended heating time while keeping the buffer and detection conditions constant (standard IHC practice). Let sections cool consistently before blocking, since uneven cooling can make staining vary across a run (standard IHC practice). Score cytoplasmic signal in hepatocytes alongside a matched negative control; hepatocytes show high staining, but the reference tissue profile carries a cross-gene specificity caution (HPA: liver hepatocytes High; HPA: Supported, targets more than one gene).
Can fixation explain weak or uneven ADH1C staining?
Target-specific fixation sensitivity is unknown from the supplied evidence, and the selected paraffin-section caption does not state a fixative (caption: A02650). Record fixative, fixation duration and tissue thickness for each specimen, then compare sections processed together before changing the antibody concentration (standard IHC practice). Use the same citrate pH 6.0 retrieval at 95–98 °C for 20 min across that comparison (page retrieval rule). Include a tissue section with an expected cytoplasmic signal to distinguish a batch-wide processing failure from a specimen-specific result (UniProt P00326: cytoplasm; HPA: liver hepatocytes High). Do not assign a particular fixation effect to ADH1C without a controlled comparison.
Does membrane or nuclear staining fit the expected ADH1C pattern?
Prioritise cytoplasmic staining when evaluating ADH1C in chromogenic sections (UniProt P00326: cytoplasm; HPA: cytosol supported). The protein has no annotated transmembrane segment, while the reported plasma membrane location is uncertain (UniProt P00326: topology; HPA subcellular: plasma membrane uncertain). Treat isolated nuclear or sharply membranous staining as a finding requiring independent validation; the additional nucleoplasmic location is uncertain and the localisation data carry a cross-gene antibody caution (HPA subcellular). Compare the suspect pattern with adjacent cytoplasmic staining, cell morphology and a negative control on the same run (standard IHC practice). A compartment mismatch alone cannot identify the source of staining.
How can I assess whether the antibody distinguishes ADH1C from related enzymes?
Check the antibody's documented immunogen or epitope against ADH1A and ADH1B sequences before assigning a stained cell specifically to ADH1C (UniProt P00326: class I ADH subunits; standard antibody validation practice). The supplied record lists 0 isoforms and no annotated domains, so an isoform-specific interpretation has no support here (UniProt P00326: isoforms and domains). ADH1C can form dimers with identical or other class I chains, but dimer membership does not establish antibody specificity (UniProt P00326: subunit). Prioritise an independently validated reagent or orthogonal expression evidence when specificity matters; the tissue reference explicitly cautions that its antibodies target proteins from more than one gene (HPA tissue: reliability description).
How should IF help resolve an ambiguous chromogenic ADH1C pattern?
Use IF as a separate validation experiment and multiplex ADH1C with a marker identifying the cell population being assessed, such as a validated hepatocyte marker when investigating liver sections (HPA: hepatocytes High; standard IF practice). Select spectrally separated fluorophores after inspecting unstained tissue for autofluorescence, and include single-label controls to assess bleed-through (standard IF practice). Because the expected signal is cytosolic, permeabilise sufficiently to admit the antibody to the cytosol, then compare with a minimally permeabilised control if apparent surface staining is the question (UniProt P00326: cytoplasm; HPA subcellular: cytosol supported). Do not transfer the paraffin IHC caption's 1:100 dilution to IF without titration (caption: A02650).
What should I check when ADH1C chromogenic staining appears widespread?
Run a no-primary control through the chromogenic workflow and inspect it for endogenous peroxidase activity or nonspecific detection-system staining (standard IHC practice). Use a peroxidase block before DAB development, then keep DAB exposure comparable across sections (standard chromogenic IHC practice). Titrate the catalog antibody around the illustrated 1:100 dilution while retaining the same retrieval and detection settings; that dilution belongs to a paraffin-embedded esophagus image, not a universal optimum (caption: A02650). Compare diffuse staining with cell-resolved cytoplasmic signal and tissue morphology (UniProt P00326: cytoplasm). Persistent signal in the no-primary control points to workflow background rather than interpretable ADH1C immunoreactivity (standard IHC practice).
How should I score ADH1C staining across tissues or treatment groups? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, and record the percentage of cytoplasm-positive cells plus staining intensity for an H-score (UniProt P00326: cytoplasm; standard IHC scoring practice). Use the same intensity scale and thresholds across batches, with a reference section in each run to monitor staining drift (standard IHC practice). Normalise positive counts or stained area to the number of eligible cells or analysed tissue area in mm², and exclude folds, necrosis and section edges by a prespecified rule (standard IHC practice). Report cell types separately: hepatocytes are high, whereas esophageal squamous epithelial cells are medium in the tissue reference (HPA: liver hepatocytes High; esophagus squamous epithelial cells Medium).
When is a positive ADH1C IHC result convincing?
A convincing result shows cell-resolved cytoplasmic staining in a plausible population, such as hepatocytes, with clean controls and preserved morphology (UniProt P00326: cytoplasm; HPA: liver hepatocytes High; standard IHC practice). Reconsider signal confined to nuclei, tissue edges or necrotic areas, or staining that persists in a no-primary control; these patterns require artefact checks (HPA subcellular: nucleoplasm uncertain; standard IHC practice). Check endogenous peroxidase contribution when DAB staining remains after primary antibody omission (standard chromogenic IHC practice). Interpret an unexpected cell type cautiously because the tissue reference reports only medium antibody–RNA consistency and warns that its antibodies target proteins from more than one gene (HPA tissue: reliability description).
Boster reagents
Best ADH1C / Alcohol dehydrogenase 1C IHC Antibodies
A02650 has real paraffin-section IHC images from human esophagus and stomach (IHC image captions) and listed Human, Mouse and Rat reactivity (catalog: A02650); no IF data are supplied (catalog: A02650).
Real IHC data
Anti-Alcohol dehydrogenase 1C ADH1C Antibody
Cat # A02650
A02650 will render with IHC images of human esophagus and stomach paraffin sections at 1:100 (IHC image captions). Its listed applications are IHC and WB, and its listed reactivity is Human, Mouse and Rat (catalog: A02650).
Which to pick: Choose A02650 for paraffin-section tissue IHC, supported by its human esophagus and stomach images (IHC image captions); the fixative is unreported (IHC image captions). There is no IF/ICC option here because A02650 has no listed IF application or IF image, and its clonality is unreported (catalog: A02650). For Mouse or Rat IHC, A02650 has listed reactivity, but the supplied tissue images show Human samples only (catalog: A02650; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P00326 (ADH1G_HUMAN, Alcohol dehydrogenase 1C).
- Human Protein Atlas. ADH1C tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ADH1C 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. ADH1C antibody validation summary (2 antibodies).
- ADH1C Down-Regulation Is a Key Hypoxia Response in Colon Epithelium. The American journal of pathology 2026 — PMC13084684.
- HCC: RNA-Sequencing in Cirrhosis. Biomolecules 2023 — PMC9855755.
- Identification of Down-Regulated ADH1C is Associated With Poor Prognosis in Colorectal Cancer Using Bioinformatics Analysis. Frontiers in molecular biosciences 2022 — PMC8921497.
- Identification of Energy Metabolism Genes for the Prediction of Survival in Hepatocellular Carcinoma. Frontiers in oncology 2020 — PMC7438573.
- PubMed PMID:3758060 — UniProt-cited evidence.
- PubMed PMID:2935875 — UniProt-cited evidence.
- PubMed PMID:1524834 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ADH1C (Alcohol dehydrogenase 1C). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02650 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Bronchus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophagus tissue; fixative not specified (datasheet A02650); 2. Fixation: Image fixative and duration unreported (datasheet A02650); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-ADH1C, 1:50-1:200 (datasheet A02650); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No isoforms annotated; mature chain spans residues 2–375 (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt P00326, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A02650, PMC13084684, PMC9855755. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ADH1C/immunohistochemistry-ihc-guide.