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GUSB / Beta-glucuronidase · IHC design guide
Design Immunohistochemistry for GUSB
Plan GUSB staining in paraffin sections using the 1:100 catalog antibody condition (datasheet A02234). Compare cytoplasmic staining in liver hepatocytes with the reported absence of staining in rectal glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GUSB 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); lysosomal protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 9 HIER, heat-mediated (datasheet A02234) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formaldehyde fixation at room temperature (selected-SKU IHC image A02234) | |
| Caveat | Staining varies by tissue; rectal glands were undetected (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | Three isoforms; epitope coverage is unresolved (UniProt) |
Section 1
Recommended GUSB IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol (datasheet A02234) with the published mouse colon protocol (PMC9140600).
| Sample | Formaldehyde-fixed, paraffin-embedded Human liver tissue (datasheet A02234) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A02234); 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 9 (datasheet A02234); 20 min, 95–100 °C (standard) |
| 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-GUSB, 1:100 (datasheet A02234) |
| Primary incubation | 1 hour at room temperature (datasheet A02234) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02234) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GUSB-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 9 retrieval for the catalog antibody (datasheet A02234); the published protocol used citrate retrieval in a pressure cooker (PMC9140600).
Section 2
What Is the Expected GUSB Staining Pattern?
GUSB is a lysosomal enzyme without a transmembrane segment (UniProt P08236). In paraffin-section IHC, expect cytoplasmic staining in many tissues, with strong staining in liver hepatocytes and several glandular cell populations (HPA tissue IHC). HPA rates the tissue staining “Approved” but reports only medium consistency with RNA expression; interpret intensity alongside the cell type and controls (HPA tissue IHC).
| Cytoplasmic staining in liver hepatocytes or adrenal, endometrial, epididymal, or gallbladder glandular cells. | These are reported high-staining populations (HPA tissue IHC). A granular appearance would fit lysosomal localization (UniProt P08236), but HPA's tissue summary establishes cytoplasmic staining, not a required punctate pattern. |
| Predominantly nuclear or sharp cell-surface staining in otherwise positive cells. | That distribution conflicts with lysosomal localization and the reported cytoplasmic tissue pattern (UniProt P08236; HPA tissue IHC). Review controls and detection before treating it as GUSB-specific staining (general IHC practice). |
| Strong staining confined to adipocytes, skeletal myocytes, or rectal glandular cells. | HPA reports GUSB as not detected in those specific cell populations (HPA tissue IHC). Consider cross-reactivity or endogenous chromogen-generating activity, and assess a no-primary control; neighboring cell types need separate interpretation (general IHC practice). |
| Uniform color across cells, stroma, and blank regions of the section. | A field-wide deposit lacks the cell-specific cytoplasmic distribution reported by HPA (HPA tissue IHC). Check the no-primary control, blocking, washing, and detection conditions for nonspecific background (general IHC practice). |
| No staining in a liver section with identifiable hepatocytes. | HPA reports high hepatocyte staining, making liver a useful positive reference (HPA tissue IHC). If the positive control also fails, first check antibody and detection performance; an unstained test section alone cannot establish absence of GUSB (general IHC practice). |
💡Expected GUSB appearanceCall a result positive when hepatocytes or the listed glandular cells show clear cytoplasmic staining, strongest in the HPA high groups (HPA tissue IHC); nuclear, crisp surface, or field-wide color is suspect given the lysosomal localization and cytoplasmic tissue profile (UniProt P08236; HPA tissue IHC).
| Cell population and tissue choice | HPA reports high staining in liver hepatocytes and selected glandular cells, medium staining in breast glands and selected brain cells, and low staining in several epithelia (HPA tissue IHC). Judge a cell population against its own reference level. |
| Subcellular interpretation | GUSB is lysosomal and has no transmembrane segment (UniProt P08236); HPA describes tissue IHC as cytoplasmic (HPA tissue IHC). Neither source requires every positive paraffin section to resolve individual lysosomes. |
| Antibody evidence | Two rabbit polyclonal antibodies, HPA036322 and HPA036323, have Approved IHC status (HPA antibodies). The supplied record does not report IHC Enhanced validation, so do not claim an enhanced level of confirmation. |
| IF/ICC Q: What pattern should appear? | A: Vesicular localization is supported in ICC-IF, with images listed for A-431 and U2OS (HPA subcellular). This is a localization cross-check, not an IHC staining-intensity standard or an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver hepatocytes and the positive control are both blank. | An assay-wide failure is plausible because hepatocytes are high in the HPA tissue profile (HPA tissue IHC). | Check primary-antibody application, detection reagents, chromogen development, and section integrity using the same control workflow (general IHC practice). |
| Test tissue is blank, but liver hepatocytes stain. | A negative result may reflect the sampled cell population: HPA reports low or undetected staining in several populations (HPA tissue IHC). | Identify the cells being scored and compare them with the corresponding HPA tissue entry; retain the successful liver control when reporting the result (HPA tissue IHC; general IHC practice). |
| Diffuse color obscures cell boundaries. | Nonspecific antibody binding or excess detection background can obscure cytoplasmic staining (general IHC practice). | Inspect a no-primary control, improve blocking and washes, and adjust antibody or detection conditions according to the assay controls (general IHC practice). |
| Color appears in a no-primary control. | Endogenous detection activity or nonspecific detection-reagent binding is possible in chromogenic IHC (general IHC practice). | Apply the appropriate endogenous-enzyme block for the detection system and verify detection reagents with control sections (general IHC practice). |
| Signal is mainly nuclear or sharply outlines the cell surface. | The distribution disagrees with the lysosomal annotation and HPA cytoplasmic tissue profile (UniProt P08236; HPA tissue IHC). | Review morphology, no-primary staining, and antibody specificity controls before scoring it as GUSB; compare with cytoplasmic staining in a known-positive cell population (general IHC practice; HPA tissue IHC). |
| Strong signal occurs in an HPA-undetected cell population. | The finding differs from HPA's cell-specific observations, though its tissue IHC has only medium consistency with RNA data (HPA tissue IHC). | Confirm the cell identity, compare a known-positive tissue and no-primary control, and describe the discrepancy rather than assigning it to GUSB from intensity alone (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing GUSB staining
A02234 · IHCImmunohistochemical analysis of A02234 on paraffin-embedded Human liver tissue. Tissue was fixed with formaldehyde at room temperature. Heat
A02234 · IHCImmunohistochemical analysis of A02234 on paraffin-embedded Human colon carcinoma tissue. Tissue was fixed with formaldehyde at room tempera
A02234-1 · IHCIHC analysis of GUSB using anti-GUSB antibody (A02234-1). GUSB was detected in a paraffin-embedded section of human liver cancer tissue. Hea
A02234-1 · IFIF analysis of GUSB using anti-GUSB antibody (A02234-1). GUSB was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169919-GUSB/tissue/cerebral+cortex#imid_9345600
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169919-GUSB/tissue/colon#imid_9345534
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169919-GUSB/tissue/liver#imid_9345543
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169919-GUSB/tissue/kidney#imid_9345544
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036322; green = GUSB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036322; green = GUSB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GUSB IHC & IF
🧪Run liver first: hepatocytes should show granular cytoplasmic staining consistent with lysosomal GUSB (HPA: High in liver hepatocytes; UniProt P08236: lysosome). Use adipose tissue as the negative, assessing adipocytes (HPA: Not detected in adipocytes); on the liver slide, cells without specific staining should show only counterstain, but no liver cell type is established here as a negative control (HPA: liver row identifies hepatocytes only).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GUSB in A-431, U2OS, with annotated localisation: Vesicles (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched isotype controls matched to the primary antibody’s host species and clonality; use GUSB-knockout material processed alongside the test sections as a biological negative (standard IHC control practice). Quench endogenous peroxidase and check liver pigment against the no-primary control before interpreting chromogen (A02234 caption: HRP polymer detection; standard liver IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC fixative is unreported (A02234 caption metadata: fixative not stated). Heat retrieval with EDTA at pH 9.0 is reported for the liver paraffin section, but a requirement for that retrieval condition is unestablished (A02234 caption: heat-induced EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier; ICC/IF images show vesicular localization in A-431 and U2OS, while liver pigment can complicate chromogenic interpretation (HPA: Vesicles supported; HPA: ICC/IF images in A-431 and U2OS; standard liver IHC practice).
HPA tissue IHC evidence for GUSB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
| Rectum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GUSB IHC Tips
Troubleshoot GUSB staining in paraffin sections by checking retrieval, cell type, cytoplasmic pattern, controls and scoring before interpreting signal.
What should I adjust when GUSB staining is weak after antigen retrieval?
Start with heat-induced retrieval in EDTA at pH 9 for paraffin sections (datasheet A02234). The selected tissue image used this retrieval, followed by A02234 at 1:100 for 1 hour at room temperature (A02234 tissue-IHC caption). If signal is weak, compare a modestly longer heating step with the current run while keeping section thickness, primary incubation and detection conditions constant (standard IHC practice). Check hepatocytes in liver as a positive reference (HPA: high in hepatocytes), and inspect tissue integrity because excessive heating can damage morphology and make apparent gains in staining difficult to interpret (standard IHC practice).
How can I troubleshoot variable GUSB staining between differently fixed paraffin blocks?
GUSB-specific sensitivity to fixation is unknown from the supplied evidence, so do not assign a weak result to a particular fixative or fixation duration (supplied GUSB evidence). Record each block’s documented fixative and processing history, then compare sections stained together with the same EDTA pH 9 retrieval and detection settings (datasheet A02234; standard IHC practice). Include a liver section containing hepatocytes as a reference in each run (HPA: high in hepatocytes; standard IHC practice). If staining differs, assess morphology, section preservation and control performance before attributing the difference to GUSB abundance (standard IHC practice).
Where should credible GUSB staining appear in paraffin tissue sections?
Expect predominantly cytoplasmic staining in tissue sections (HPA: cytoplasmic expression in most tissues), consistent with a lysosomal protein (UniProt P08236: lysosome). A fine punctate pattern can support a vesicular interpretation, although chromogenic resolution may merge small structures into broader cytoplasmic signal (HPA subcellular: vesicles supported; standard IHC practice). GUSB has no annotated transmembrane segment, so isolated sharp plasma-membrane outlining needs scrutiny (UniProt P08236 topology). Compare the distribution within hepatocytes, which show high tissue-IHC expression, against adjacent morphology and a no-primary control before accepting an unexpected nuclear or surface-dominant pattern (HPA: high in hepatocytes; standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent GUSB staining?
GUSB has 3 annotated isoforms, but the supplied evidence does not map A02234’s epitope to any of them (UniProt P08236: isoforms 1–3; supplied A02234 evidence). Its precursor has a signal peptide at residues 1–22 and a documented chain at 23–651, so an epitope assignment would need sequence-level support (UniProt P08236 processing). Four annotated glycosylation sites at residues 173, 272, 420 and 631 make epitope accessibility a plausible question, not an established staining effect (UniProt P08236 glycosylation). Compare retrieval conditions and positive-control morphology first; request the antibody’s mapped epitope before claiming isoform-specific staining (standard IHC practice).
How should I assess GUSB by IF alongside this chromogenic IHC guide?
Use this as an IF troubleshooting check, with the antibody’s IF performance evaluated separately from the paraffin IHC image (A02234 tissue-IHC caption; standard IF practice). Multiplex GUSB with a marker identifying the expected cell population, such as hepatocytes in liver, and check whether puncta occupy the expected cells (HPA: high in hepatocytes; HPA subcellular: vesicles supported). Choose a fluorophore channel with low tissue autofluorescence and include single-color controls for bleed-through (standard IF practice). Because GUSB is lysosomal and lacks a transmembrane segment, permeabilise fixed cells to access an intracellular epitope, then confirm that the chosen treatment preserves vesicle morphology (UniProt P08236 localisation and topology; standard IF practice).
What causes diffuse or granular background in GUSB chromogenic staining?
First distinguish staining within cells from diffuse deposit on folds, edges or damaged tissue (standard IHC practice). GUSB is a lysosomal protein with supported vesicular localisation, so intracellular granularity can be credible when it follows intact cell boundaries (UniProt P08236: lysosome; HPA subcellular: vesicles supported). Run a no-primary control and a peroxidase block before HRP detection to assess nonspecific reagent deposition and endogenous peroxidase activity (standard IHC practice). If background persists, titrate the primary around the caption’s 1:100 starting dilution and shorten chromogen development while preserving the same positive reference and counterstain (A02234 tissue-IHC caption; standard IHC practice).
How should I score GUSB staining across cells and tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive cells and staining intensity as an H-score or measure positive-cell density per mm² (standard IHC practice). Keep retrieval, antibody incubation and chromogen development consistent; the selected image used EDTA pH 9 and primary antibody at 1:100 for 1 hour (A02234 tissue-IHC caption). Normalise counts to evaluable cells or intact tissue area, and exclude folds, necrosis and section edges using a prespecified rule (standard IHC practice). Report hepatocytes separately from other liver cells because the HPA tissue reference identifies high expression specifically in hepatocytes (HPA: high in hepatocytes).
How can I separate genuine GUSB signal from staining artefacts?
A convincing result shows intracellular cytoplasmic staining in intact cells, potentially with vesicular granularity (HPA: cytoplasmic expression in most tissues; HPA subcellular: vesicles supported). In liver, check hepatocytes as the expected high-expression population, and treat signal confined to an unexpected cell population as a finding requiring independent confirmation (HPA: high in hepatocytes; standard IHC practice). Reject patterns limited to section edges, folds or necrotic areas, and investigate dominant nuclear or plasma-membrane staining against the lysosomal assignment and lack of a transmembrane segment (standard IHC practice; UniProt P08236 localisation and topology). Compare no-primary and peroxidase-block controls before interpreting chromogen deposits, since endogenous enzyme activity can mimic antibody-dependent signal (standard IHC practice).
Boster reagents
Best GUSB / Beta-glucuronidase IHC Antibodies
Real IHC data cover human paraffin liver, colon carcinoma, and liver cancer (catalog IHC captions); IF data cover A431 cells (A02234-1 IF caption). A02234 also lists mouse reactivity (catalog reactivity).
Real IHC data
Anti-GUSB Antibody (Center)
Cat # A02234
Real IHC data
Anti-beta glucuronidase/GUSB Antibody ®
Cat # A02234-1
A02234 shows IHC on paraffin-embedded human liver and colon carcinoma, with formaldehyde fixation and EDTA pH 9.0 retrieval (A02234 IHC captions). A02234-1 shows IHC on paraffin-embedded human liver cancer with EDTA pH 8.0 retrieval, plus IF in A431 cells (A02234-1 IHC and IF captions).
Which to pick: For tissue IHC, A02234 has documented human paraffin-section staining after formaldehyde fixation and EDTA pH 9.0 retrieval at 1:100 (A02234 IHC captions). For IF/ICC, A02234-1 lists both applications and shows A431-cell IF at 2 μg/mL; its paraffin liver cancer IHC caption reports EDTA pH 8.0 retrieval but does not report a fixative (A02234-1 applications; A02234-1 IF and IHC captions). For mouse work, A02234 lists mouse reactivity and IHC-P, while its IHC images show human samples (A02234 catalog reactivity and applications; A02234 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P08236 (BGLR_HUMAN, Beta-glucuronidase).
- Human Protein Atlas. GUSB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GUSB subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. GUSB antibody validation summary (2 antibodies).
- Decreased Tissue Omega-6/Omega-3 Fatty Acid Ratio Prevents Chemotherapy-Induced Gastrointestinal Toxicity Associated with Alterations of Gut Microbiome. International journal of molecular sciences 2022 — PMC9140600.
- Evaluation of AAV-mediated Gene Therapy for Central Nervous System Disease in Canine Mucopolysaccharidosis VII. Molecular therapy : the journal of the American Society of Gene Therapy 2016 — PMC4817811.
- The effect of neonatal gene therapy on skeletal manifestations in mucopolysaccharidosis VII dogs after a decade. Molecular genetics and metabolism 2013 — PMC3690974.
- New genetic prognostic biomarkers in primary central nervous system lymphoma (PCNSL). Brain and behavior 2021 — PMC8035458.
- PubMed PMID:3468507 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GUSB (Beta-glucuronidase). Expected localisation Cytoplasmic staining (HPA tissue IHC); lysosomal protein (UniProt). Antigen retrieval: EDTA pH 9 HIER, heat-mediated (datasheet A02234). Validated IHC antibody: A02234 at 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded Human liver tissue (datasheet A02234); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A02234); 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 9 (datasheet A02234); 20 min, 95–100 °C (standard); 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-GUSB, 1:100 (datasheet A02234); 9. Primary incubation: 1 hour at room temperature (datasheet A02234). Watch for: Fixation: Formaldehyde fixation at room temperature (selected-SKU IHC image A02234); Caveat: Staining varies by tissue; rectal glands were undetected (HPA tissue IHC); Isoform / epitope: Three isoforms; epitope coverage is unresolved (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P08236, HPA tissue IHC · Approved, HPA ICC-IF · vesicles., Datasheet A02234, PMC9140600. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GUSB/immunohistochemistry-ihc-guide.