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HGD / Homogentisate 1,2-dioxygenase · IHC design guide
Design Immunohistochemistry for HGD
Plan chromogenic HGD IHC in paraffin sections using liver hepatocytes as a positive control (HPA tissue IHC). Start catalog antibody A01909-1 at 0.5–1 μg/ml (datasheet A01909-1) and compare with adipocytes, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HGD 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 in a subset of tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in hepatocytes and glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01909-1) | |
| Positive control | Endometrium+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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Liver endogenous peroxidase can mimic DAB staining (standard IHC practice) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–445 chain (UniProt) |
Section 1
Recommended HGD IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published HGD IHC method using mouse tissues (PMC13200543).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A01909-1) |
| Fixation | Image fixative and duration unreported (datasheet A01909-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01909-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01909-1) |
| Primary antibody | Rabbit anti-HGD, 0.5-1μg/ml recommended; image 2μg/ml (datasheet A01909-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01909-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01909-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HGD-positive staining in ciliated epithelial cells of endometrium (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a subset of tissues including liver, kidney, gallbladder, gastrointestinal tract, prostate and thyroid gland. No signal in the no-primary control. |
💡Decision noteTry citrate pH 6 heat retrieval first (datasheet A01909-1); the published method does not specify retrieval (PMC13200543).
Section 2
What Is the Expected HGD Staining Pattern?
For paraffin-section IHC, expect HGD staining in the cytoplasm of selected epithelial cells, especially hepatocytes and gallbladder glandular cells (HPA: tissue IHC, High). Kidney proximal tubule cell bodies and prostate glandular cells show Medium staining (HPA: tissue IHC). The tissue pattern has Enhanced reliability (HPA: tissue IHC). HGD has no annotated transmembrane segment (UniProt Q93099 topology); its Golgi assignment comes from ICC-IF (HPA: subcellular ICC-IF).
| Strong cytoplasmic staining in hepatocytes, with staining concentrated in the expected cells. | This matches a High tissue-IHC result (HPA: liver hepatocytes, High). Judge the pattern by cell identity and compartment as well as intensity; staining throughout unrelated cells does not reproduce the reported distribution (HPA: tissue IHC profile). |
| Predominantly nuclear, membrane-rim or extracellular staining in an otherwise positive section. | Treat this as a pattern mismatch: HPA reports cytoplasmic tissue staining (HPA: tissue IHC profile), and UniProt annotates no transmembrane segment (UniProt Q93099 topology). Check controls and detection background before interpreting such staining as HGD. |
| Strong staining in adipocytes or bone-marrow hematopoietic cells. | These cells are reported as Not detected (HPA: adipose tissue adipocytes; bone marrow hematopoietic cells). Review the stained cell type and the no-primary control; cross-reactivity or endogenous detection activity may explain unexpected signal (general IHC practice). |
| Diffuse color across cells, stroma and the section background. | A field-wide deposit obscures the selective cytoplasmic pattern (HPA: tissue IHC profile). Compare a no-primary control and review blocking, washes and detection chemistry for nonspecific or endogenous signal (general IHC practice). |
| No detectable staining in a well-preserved liver section. | Hepatocytes are a High reference population (HPA: liver hepatocytes, High). A blank result warrants review of the IHC-validated antibody, retrieval and detection controls (general IHC practice); this section has no evidence for an HGD-specific fixation effect. |
💡Expected HGD appearanceCall a result positive when selected cells show cytoplasmic staining, ideally High in hepatocytes or gallbladder glandular cells (HPA: tissue IHC); widespread nuclear or unrelated-cell staining is suspect against that pattern (HPA: tissue IHC profile).
| Choice of reference tissue and cell | Liver hepatocytes and gallbladder glandular cells provide High examples; kidney proximal tubules and prostate glandular cells are Medium (HPA: tissue IHC). Score the named cells, since whole-section intensity can obscure a selective result. |
| Strength of tissue-pattern evidence | Tissue IHC is rated Enhanced for consistency with RNA expression (HPA: tissue IHC reliability). Two listed antibodies have Enhanced IHC validation, HPA047374 and HPA052359 (HPA: antibodies); this supports the reported pattern without validating every staining condition. |
| Cellular location and assay | HPA describes cytoplasmic staining in tissue IHC, while its approved Golgi location comes from ICC-IF (HPA: tissue IHC; subcellular ICC-IF). The ICC-IF result alone does not require Golgi-resolved chromogenic staining in paraffin sections. |
| Protein architecture | HGD is a single annotated chain, residues 1–445, with no signal peptide, propeptide or transmembrane segment (UniProt Q93099 processing and topology). Those annotations provide no basis for expecting a shed or membrane-anchored IHC pattern. |
| IF/ICC Q: What location should I compare? | A: Compare a cellular IF/ICC result with the approved Golgi assignment (HPA: subcellular ICC-IF). That evidence is assay-specific; the paraffin-section comparison remains the cytoplasmic tissue-IHC profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver control is blank while the counterstain and tissue morphology are visible. | Hepatocytes normally provide a High signal (HPA: liver hepatocytes, High); a blank control can reflect an assay or detection failure (general IHC practice). | Check the IHC-validated antibody, retrieval, primary incubation and detection controls against the run record (general IHC practice). Do not infer HGD-specific fixation sensitivity from this result. |
| Kidney appears weaker than liver. | That difference is consistent with the reported levels: proximal tubule cell bodies are Medium and hepatocytes are High (HPA: kidney; liver tissue IHC). | Score proximal tubule cell bodies separately and retain a liver reference in the comparison (HPA: kidney; liver tissue IHC). Investigate only if the expected cell-restricted signal disappears. |
| Signal is confined to nuclei or outlines cell membranes. | The compartment conflicts with cytoplasmic tissue staining (HPA: tissue IHC profile); HGD has no annotated transmembrane segment (UniProt Q93099 topology). | Inspect the no-primary control, stain distribution and cell morphology; reassess staining specificity before assigning the pattern to HGD (general IHC practice). |
| Strong color appears in adipocytes or bone-marrow hematopoietic cells. | Both populations are listed as Not detected (HPA: adipose tissue; bone marrow tissue IHC). Cross-reactivity or endogenous detection activity is a possible IHC explanation, not an established HGD finding. | Compare a no-primary control, review blocking and detection chemistry, and confirm the cell identity before scoring a positive result (general IHC practice). |
| Most cells and extracellular areas carry a similar faint stain. | Diffuse deposit can mask the selective cytoplasmic profile (HPA: tissue IHC profile); nonspecific staining or background detection is possible (general IHC practice). | Compare background with the no-primary control, then review blocking, wash stringency and detection steps (general IHC practice). Score only staining that remains cell-restricted. |
| An IF/ICC image looks Golgi-localized, but paraffin IHC looks broadly cytoplasmic. | The Golgi assignment is from ICC-IF, while the broader cytoplasmic description is from tissue IHC (HPA: subcellular ICC-IF; tissue IHC). | Interpret each image against its own assay's reference pattern. For paraffin IHC, prioritize the reported stained cell types and cytoplasmic distribution (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing HGD staining
A01909-1 · IHCIHC analysis of HGD using anti-HGD antibody (A01909-1). HGD was detected in paraffin-embedded section of human liver cancer tissue. Heat med
A01909-1 · IHCIHC analysis of HGD using anti-HGD antibody (A01909-1). HGD was detected in paraffin-embedded section of human prostatic cancer tissue. Heat
A01909-1 · IFIF analysis of HGD using anti-HGD antibody (A01909-1). HGD was detected in immunocytochemical section of CACO-2 cells. Enzyme antigen retrie
M01909-1 · IHCIHC analysis of HGD using anti-HGD antibody (M01909-1). HGD was detected in a paraffin-embedded section of human liver cancer tissue. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000113924-HGD/tissue/cerebral+cortex#imid_12896735
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000113924-HGD/tissue/colon#imid_12896645
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000113924-HGD/tissue/liver#imid_12896646
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000113924-HGD/tissue/gallbladder#imid_12896622
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA052359; green = HGD, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA052359; green = HGD, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HGD IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in hepatocytes). Use adipose tissue as the negative tissue, where adipocytes were not detected (HPA: Not detected in adipocytes); on the liver slide, assess background in adjacent non-hepatocyte cells without treating them as validated HGD-negative cells.
Positive control tissue: Endometrium (Ciliated epithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HGD in Hep-G2, RPTEC/TERT1, U2OS, with annotated localisation: Golgi apparatus (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched rabbit IgG isotype control (catalog antibody caption: rabbit anti-HGD), and HGD-knockout tissue or a peptide-block control if available. For liver, quench endogenous peroxidase and control for endogenous biotin when using the caption’s biotin-based DAB detection (catalog antibody caption: SABC with DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01909-1 tissue-IHC caption does not state the fixative (catalog antibody caption: fixative not stated). The demonstrated paraffin-section workflow uses heat retrieval in citrate buffer at pH 6 for 20 minutes; retrieval dependence and frozen-section performance are unreported (catalog antibody caption: citrate retrieval). IF feasibility in tissue is unreported, although ICC-IF images exist for Hep-G2, RPTEC/TERT1 and U2OS (HPA: subcellular); in liver, check endogenous biotin and peroxidase background with the caption’s SABC–DAB method (catalog antibody caption: SABC with DAB).
HPA tissue IHC evidence for HGD
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 |
|---|---|---|---|---|
| Endometrium | Ciliated epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | Proximal tubules (cell body) | 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HGD IHC Tips
Troubleshoot HGD staining in paraffin sections by checking retrieval, cell type, localisation and controls alongside the chromogenic signal.
What retrieval should I try first when HGD staining is weak?
Start with heat-mediated antigen retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01909-1). The selected paraffin-section image used that retrieval before overnight incubation with 2 µg/ml catalog antibody at 4°C (datasheet A01909-1). If signal remains weak, compare a carefully controlled longer heating interval on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Score hepatocytes or kidney proximal tubules against a matched negative control, because those cells have documented HGD staining; avoid judging retrieval from uniformly unstained tissue alone (HPA: tissue IHC).
How should I investigate variable HGD staining across differently fixed blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, and the supplied HPA tissue profile does not establish a fixation response (datasheet A01909-1; HPA: tissue IHC). Record each block’s fixative, time before fixation and fixation duration, then compare sections processed with the same citrate pH 6 retrieval (standard IHC practice; datasheet A01909-1). Use the same antibody concentration and detection run for that comparison to limit technical variation (standard IHC practice). A difference between blocks identifies a condition worth investigating, but it does not by itself establish that fixation altered the HGD epitope (standard IHC practice).
Should HGD appear diffusely cytoplasmic or concentrated near the Golgi?
In tissue IHC, HGD is reported as cytoplasmic in a subset of tissues, including liver and kidney (HPA: tissue IHC). Separate ICC/IF evidence assigns HGD an approved Golgi apparatus location, so a concentrated intracellular pattern can also be compatible with the supplied evidence (HPA: subcellular). HGD has no annotated transmembrane segment, while UniProt supplies no subcellular location annotation; neither fact establishes a membrane-side staining pattern in paraffin sections (UniProt Q93099: topology and subcellular annotation). Compare the stained cells with a tissue-appropriate positive control and inspect nuclear or surface-restricted signal against negative controls before interpreting it as HGD (HPA: tissue IHC; standard IHC practice).
Could an isoform or modified residue explain inconsistent HGD immunostaining?
The supplied record lists 0 isoforms and one chain spanning residues 1–445, so it offers no documented alternative isoform to explain different IHC patterns (UniProt Q93099: processing and isoforms). It lists acetylation at residue 98 and succinylation at residue 414, but provides no antibody epitope map or evidence that either modification changes binding (UniProt Q93099: modified residues; datasheet A01909-1). Treat epitope masking as a hypothesis and compare adjacent sections under the documented citrate pH 6 retrieval first (datasheet A01909-1; standard IHC practice). If staining remains inconsistent, use an independently validated antibody or orthogonal expression evidence to test specificity (standard IHC practice).
How can IF help assess an ambiguous chromogenic HGD pattern?
Use IF as a separate localisation check, since HPA reports an approved Golgi apparatus location from ICC/IF while tissue IHC reports cytoplasmic staining (HPA: subcellular; HPA: tissue IHC). Multiplex HGD with a marker identifying the expected cell type, such as hepatocytes in liver, and compare cell identity rather than fluorescence intensity alone (HPA: tissue IHC; standard IF practice). Select spectrally separated fluorophores and favor a red or far-red HGD channel when tissue autofluorescence is prominent, with single-channel and no-primary controls (standard IF practice). Because the antibody epitope is unspecified, test mild permeabilisation for intracellular access without assuming a membrane-facing epitope, and assess whether the Golgi pattern is preserved (UniProt Q93099: topology; HPA: subcellular; standard IF practice).
How do I distinguish background DAB deposition from genuine HGD staining?
The selected image used a biotinylated secondary antibody, streptavidin-biotin detection and DAB, so check background at each detection stage with a no-primary section (datasheet A01909-1; standard IHC practice). Include a peroxidase block and verify that the no-primary control remains clean before attributing brown signal to HGD (standard IHC practice). If diffuse colour persists, reassess blocking, washes, antibody concentration and development time while retaining the documented 2 µg/ml starting concentration as a comparison point (datasheet A01909-1; standard IHC practice). Give greater weight to staining in expected cells, such as hepatocytes, than to deposits without cellular localisation (HPA: tissue IHC; standard IHC practice).
What should I measure when comparing HGD IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population before scoring: hepatocytes show high HGD staining, whereas kidney proximal tubule cell bodies show medium staining in the supplied tissue profile (HPA: tissue IHC). Within that population, report the percentage of positive cells and an intensity-weighted H-score from 0–300, using a prespecified threshold and the same counterstain and imaging conditions (standard IHC practice). Normalise positive counts to all eligible cells rather than total section area when tissue composition differs (standard IHC practice). If measuring stained area or density per mm², report the analysed viable tissue area and exclude folds, necrosis and edges using the same rules across sections (standard IHC practice).
Which staining features make an HGD-positive result convincing?
A convincing result places intracellular signal in an expected cell population, such as liver hepatocytes, and reproduces it across intact tissue away from section edges (HPA: tissue IHC; standard IHC practice). Tissue IHC describes cytoplasmic HGD, while ICC/IF supports a Golgi location; isolated nuclear or cell-surface staining therefore needs additional validation (HPA: tissue IHC; HPA: subcellular). Discount signal confined to necrotic regions or edges, and check no-primary and peroxidase-block controls for chromogen generated independently of primary-antibody binding (standard IHC practice). Compare against a low-signal reference such as adipocytes in adipose tissue, reported as not detected, while allowing for differences in tissue processing (HPA: tissue IHC; standard IHC practice).
Boster reagents
Best HGD / Homogentisate 1,2-dioxygenase IHC Antibodies
Anti-HGD antibodies have IHC images from paraffin-embedded human cancer sections (catalog IHC captions); A01909-1 also has an IF/ICC image from CACO-2 cells and lists human, mouse and rat reactivity (catalog IF caption; reactivity).
Real IHC data
Anti-HGD Antibody ®
Cat # A01909-1
Real IHC data
Anti-HGD Antibody ® (monoclonal, 2F11E1)
Cat # M01909-1
A01909-1 has IHC images from paraffin-embedded human liver and prostatic cancer sections and an IF/ICC image from CACO-2 cells (A01909-1 image captions). M01909-1 has an IHC image from a paraffin-embedded human liver cancer section (M01909-1 IHC caption).
Which to pick: For human tissue IHC, M01909-1 is a mouse monoclonal with a paraffin-section image using EDTA retrieval; A01909-1 is a rabbit antibody with paraffin-section images using citrate retrieval (catalog antibody details; respective IHC captions). For IF/ICC, choose A01909-1 because those applications are listed and its IF image shows CACO-2 cells (A01909-1 applications; IF caption). For cross-species IHC, A01909-1 lists human, mouse and rat reactivity, while M01909-1 lists human only; the fixative is unreported in both paraffin-section IHC captions (catalog reactivity; respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q93099 (HGD_HUMAN, Homogentisate 1,2-dioxygenase).
- Human Protein Atlas. HGD tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HGD subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. HGD antibody validation summary (2 antibodies).
- Demarcated redness associated with increased vascular density/size: a useful marker of flat-type dysplasia in patients with ulcerative colitis. Endoscopy international open 2021 — PMC8041573.
- Specific knockout of kidney homogentisate 1,2-dioxygenase reveals that local metabolism of tyrosine and homogentisic acid is negligible in alkaptonuria. Human molecular genetics 2026 — PMC13200543.
- Expression analysis of Barrett's esophagus-associated high-grade dysplasia in laser capture microdissected archival tissue. Clinical cancer research : an official journal of the American Association for Cancer Research 2008 — PMC2701739.
- Expression of ATF6 as a marker of pre-cancerous atypical change in ulcerative colitis-associated colorectal cancer: a potential role in the management of dysplasia. Journal of gastroenterology 2018 — PMC5910497.
- PubMed PMID:8782815 — UniProt-cited evidence.
- PubMed PMID:9244427 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HGD (Homogentisate 1,2-dioxygenase). Expected localisation Cytoplasmic staining in a subset of tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01909-1). Validated IHC antibody: A01909-1 at 0.5-1μg/ml recommended; image 2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Endometrium (Ciliated epithelial 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 A01909-1); 2. Fixation: Image fixative and duration unreported (datasheet A01909-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6 , 20 min (datasheet A01909-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01909-1); 8. Primary antibody: Rabbit anti-HGD, 0.5-1μg/ml recommended; image 2μg/ml (datasheet A01909-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01909-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Liver endogenous peroxidase can mimic DAB staining (standard IHC practice); Isoform / epitope: No annotated isoforms; one 1–445 chain (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt Q93099, HPA tissue IHC · Enhanced, HPA ICC-IF · Golgi apparatus., Datasheet A01909-1, PMC13200543. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HGD/immunohistochemistry-ihc-guide.