This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
GUSB / Beta-glucuronidase · IHC design guide
Design Immunohistochemistry for GUSB
Plan chromogenic GUSB IHC in paraffin sections using liver hepatocytes as a high-staining reference (HPA tissue IHC). Expect cytoplasmic tissue staining while interpreting GUSB as a lysosomal protein (HPA tissue IHC; UniProt).
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-fixed sections are documented (selected-SKU IHC image A02234); keep fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Endogenous peroxidase can mimic HRP/DAB staining (standard IHC practice) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; mature chain begins at residue 23 (UniProt) |
Section 1
Recommended GUSB IHC & IF Protocols
The catalog antibody uses EDTA pH 9 heat retrieval (datasheet A02234). The published mouse colon protocol below uses citrate retrieval (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 EDTA pH 9 heat retrieval for the catalog antibody (datasheet A02234). Use citrate retrieval when reproducing the published mouse colon protocol (PMC9140600).
Section 2
What Is the Expected GUSB Staining Pattern?
GUSB is a lysosomal enzyme without a transmembrane segment (UniProt P08236: location, topology). In paraffin IHC, expect cytoplasmic staining in most tissues, especially glandular cells in adrenal gland, endometrium, epididymis and gallbladder, and hepatocytes in liver (HPA: tissue IHC). HPA rates its tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA: reliability).
| Cytoplasmic staining in adrenal gland, endometrial, epididymal or gallbladder glandular cells, or liver hepatocytes. | This fits HPA's High staining in those cells and its broader cytoplasmic profile (HPA: tissue IHC). A chromogenic slide need not resolve individual lysosomes to be consistent with lysosomal GUSB (UniProt P08236: location). |
| Predominantly nuclear, membranous or extracellular signal, with little cytoplasmic staining. | That distribution conflicts with the reported cytoplasmic tissue pattern and lysosomal location (HPA: tissue IHC; UniProt P08236: location). Check staining specificity and detection background before calling it GUSB; tissue IHC alone cannot identify the source of the signal. |
| Strong staining in a cell population HPA lists as Not detected, such as adipocytes or skeletal myocytes. | Treat it as unexpected for that cell type (HPA: adipocytes and myocytes, Not detected). Consider nonspecific antibody binding or endogenous detection activity (general IHC practice). An HPA negative is a reference pattern, not proof that every specimen must be negative. |
| Widespread, fairly uniform chromogen obscures cell borders and expected tissue differences. | The diffuse background makes the reported cell specific pattern difficult to judge (HPA: tissue IHC). Nonspecific binding or endogenous detection activity can produce background (general IHC practice); evaluate controls before assigning it to GUSB. |
| No convincing cytoplasmic signal in a well preserved liver section or another HPA High tissue. | This is inconsistent with the HPA reference pattern for hepatocytes or the specified glandular cells (HPA: High in liver, adrenal gland, endometrium, epididymis and gallbladder). Review the IHC run and controls before interpreting the sample as GUSB negative (general IHC practice). |
💡Expected GUSB appearanceCall a result positive when cytoplasmic chromogen is clear in HPA High glandular cells or hepatocytes; predominantly nuclear signal or strong staining in HPA Not detected cells warrants review (HPA: tissue IHC; UniProt P08236: lysosome).
| Compartment and resolution | UniProt places GUSB in lysosomes, while HPA tissue IHC describes cytoplasmic expression (UniProt P08236: location; HPA: tissue IHC). Judge chromogenic sections by a cytoplasmic pattern; discrete vesicles are documented in HPA ICC-IF, where they can be resolved more readily (HPA: subcellular). |
| Choice of reference tissue and cell | Adrenal gland, endometrium, epididymis and gallbladder glandular cells, plus liver hepatocytes, are HPA High examples (HPA: tissue IHC). Breast glandular cells and caudate glia are Medium; several other cell types are Low or Not detected, so score the specified cell population rather than the whole section (HPA: tissue IHC). |
| Strength of tissue evidence | HPA labels the tissue IHC profile Approved and describes medium consistency with RNA data; RNA tissue specificity is Low (HPA: reliability, RNA specificity). Use the listed staining levels as reference observations, with that stated validation limit, rather than absolute thresholds for a new specimen. |
| Processing, glycosylation and isoforms | GUSB has a signal peptide at residues 1–22, a mature chain at 23–651, four annotated glycosylation sites and three isoforms (UniProt P08236: processing, glycosylation, isoforms). Neither source supplies the antibody epitope or isoform coverage, so these annotations cannot predict staining intensity or retrieval needs. |
| Topology and protocol limits | No transmembrane segment is annotated (UniProt P08236: topology). That supports checking an apparent membrane only pattern, but it does not specify antigen retrieval or fixation sensitivity; no target specific fixation effect is supplied (UniProt P08236: topology; HPA: tissue IHC scope). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA High tissue shows no staining. | The result conflicts with the reference cell pattern; the supplied sources do not establish which workflow step failed (HPA: High in liver and listed glandular cells). | Confirm that the intended cell population is present and that run controls worked. Review retrieval, primary antibody use and detection against the selected IHC-P protocol (general IHC practice); no dilution or retrieval setting is supplied here. |
| Signal is mainly nuclear or outlines cell membranes. | This conflicts with cytoplasmic tissue staining and lysosomal localization (HPA: tissue IHC; UniProt P08236: location). | Compare a HPA High reference tissue and the negative control in the same run. Reassess specificity and background before scoring the unexpected compartment as positive (general IHC practice). |
| Adipocytes, skeletal myocytes or rectal glandular cells stain strongly. | HPA reports those cell types as Not detected (HPA: tissue IHC); nonspecific binding or endogenous detection activity is possible (general IHC practice). | Check the exact cell identity, then examine a negative control and detection controls. Report the disagreement with HPA rather than treating one reference negative as an absolute biological rule. |
| Chromogen is diffuse across cells or the entire section. | Diffuse background can conceal the cell pattern; nonspecific binding or endogenous detection activity are general IHC possibilities (general IHC practice). | Review blocking, washes, primary antibody concentration and detection controls using the chosen IHC-P workflow (general IHC practice). Reassess the HPA expected cell pattern only after background is controlled (HPA: tissue IHC). |
| Staining differs between HPA High and Low tissues less than expected. | HPA levels describe observations in specified cell populations; its Approved profile has medium consistency with RNA data (HPA: tissue IHC, reliability). | Score the named cell types separately and compare sections processed in the same run (general IHC practice). Record the observed difference without converting HPA categories into a quantitative cutoff. |
| IF/ICC Q: Must a separate fluorescence protocol be inferred from the IHC result? | HPA reports vesicular ICC-IF localization and lists HPA036322 as ICC Supported (HPA: subcellular, antibodies); its tissue IHC profile is cytoplasmic (HPA: tissue IHC). | A: Use vesicular localization as the IF/ICC pattern check (HPA: subcellular). Consult the separate IF/ICC guide for that application; the IHC tissue pattern does not specify an IF/ICC protocol. |
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 stain (HPA: High in hepatocytes). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); within the liver section, hepatocyte nuclei and extracellular spaces should lack specific staining (UniProt P08236: lysosome).
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 a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, clonality, and concentration; and GUSB-knockout tissue or cells as a biological negative. For liver IHC with HRP detection (A02234 caption: HRP polymer), block endogenous peroxidase and assess residual background.
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported. The A02234 liver paraffin-section caption describes formaldehyde fixation at room temperature and heat retrieval in EDTA at pH 9.0, but gives no fixation duration or comparison establishing retrieval dependency. HPA ICC-IF shows vesicular localization in A-431 and U2OS cells (HPA: Vesicles supported); the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC.
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 section IHC using the catalog antibody’s documented retrieval conditions, expected lysosomal localisation, and tissue staining controls.
Which retrieval conditions should I try first for weak GUSB staining?
Start with heat mediated retrieval in EDTA at pH 9 (datasheet A02234). The catalog antibody’s paraffin section image used this retrieval, followed by primary antibody at 1:100 for 1 hour at room temperature (caption A02234). If signal is weak, compare retrieval heating and cooling conditions on adjacent sections while keeping detection and development constant (standard IHC practice). Include liver hepatocytes as a positive reference and adipocytes as a low signal comparator (HPA: High in liver hepatocytes; Not detected in adipocytes). Record tissue preservation alongside staining intensity, since excessive retrieval can damage section morphology (standard IHC practice).
Could fixation explain weak or uneven GUSB staining?
GUSB specific sensitivity to fixation is unknown from the supplied evidence; evaluate fixation as a possible source of variation without assuming a particular effect (supplied evidence; standard IHC practice). Compare sections with documented processing histories under the same EDTA pH 9 retrieval and 1:100 primary conditions (datasheet A02234; caption A02234). Check whether poor nuclear detail, detached tissue, or broad loss of staining accompanies the weak signal (standard IHC practice). Use liver hepatocytes as a reference for expected staining, while recognising that their HPA staining pattern does not establish fixation tolerance (HPA: High in liver hepatocytes).
Where should a convincing GUSB chromogenic signal appear?
Expect predominantly cytoplasmic staining with a granular, vesicular distribution consistent with a lysosomal enzyme (HPA: Cytoplasmic expression in most tissues; HPA subcellular: Vesicles; UniProt P08236: Lysosome). Assess localisation within intact cells rather than treating diffuse staining across a section as equally persuasive (standard IHC practice). GUSB has a signal peptide at residues 1–22 and no transmembrane segment, so a sharp continuous plasma membrane outline is unexpected (UniProt P08236 topology and processing). Compare the suspicious pattern with hepatocytes, where staining is reported as High, and adipocytes, where it is Not detected (HPA tissue IHC). Counterstain and inspect morphology before assigning staining to a cell population (standard IHC practice).
Can this IHC antibody distinguish GUSB isoforms or glycosylation states?
Do not assign an isoform from chromogenic staining alone: the record lists 3 GUSB isoforms, but the supplied antibody evidence does not map its epitope to any one of them (UniProt P08236 isoforms; supplied antibody evidence). The annotated mature chain spans residues 23–651, following a 1–22 signal peptide (UniProt P08236 processing). Four glycosylation sites are annotated at 173, 272, 420, and 631; their presence does not establish whether this antibody’s IHC epitope is affected (UniProt P08236 glycosylation; supplied antibody evidence). For an isoform specific claim, obtain epitope mapping and orthogonal isoform evidence before interpreting staining differences (standard IHC practice).
How should I cross check the IHC pattern by immunofluorescence?
Use IF as a spatial cross check of the paraffin section IHC result, with a marker for the cell population being assessed; liver hepatocytes are a documented High staining reference (HPA tissue IHC; standard IF practice). Choose fluorophores in channels with low measured tissue autofluorescence and include a no primary control for each imaging setup (standard IF practice). Because GUSB is lysosomal and lacks a transmembrane segment, test permeabilisation sufficient to expose an intracellular epitope while preserving vesicle morphology (UniProt P08236 localisation and topology; standard IF practice). Compare punctate signal with the supported vesicular IF localisation, but do not transfer the IHC antibody’s 1:100 dilution to IF without validation (HPA subcellular; caption A02234).
How can I separate GUSB staining from chromogenic background?
Run a no primary section through the same polymer and chromogen steps to reveal detection related background (standard IHC practice). Include a peroxidase block before HRP detection, then compare development times across matched sections (standard IHC practice; caption A02234: HRP polymer detection). Judge whether signal follows intact cell cytoplasm and vesicle rich regions rather than folds, section edges, or damaged tissue (HPA: Cytoplasmic expression; HPA subcellular: Vesicles; standard IHC practice). Hepatocytes offer a High staining reference, whereas adipocytes are reported as Not detected; neither reference alone replaces the no primary control (HPA tissue IHC; standard IHC practice).
How should I quantify GUSB staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then use an H-score or report the percentage of positive cells with intensity categories (standard IHC practice). If counting stained cells, normalise to all evaluable cells of that population; for area based counts, report cells per mm² of viable tissue (standard IHC practice). Keep retrieval at EDTA pH 9, the documented 1:100 primary condition, and chromogen development consistent across the comparison (datasheet A02234; caption A02234; standard IHC practice). Score hepatocytes separately from other liver cells because HPA identifies hepatocytes specifically as High, and exclude folds or necrotic areas by a predefined rule (HPA tissue IHC; standard IHC practice).
What would make an apparent GUSB positive result unconvincing?
A convincing result should localise to intact cell cytoplasm with a vesicular pattern consistent with lysosomal GUSB (HPA: Cytoplasmic expression; HPA subcellular: Vesicles; UniProt P08236: Lysosome). Treat isolated nuclear or continuous membrane staining cautiously because those patterns conflict with the supplied localisation and topology (UniProt P08236 localisation and topology; standard IHC practice). Recheck apparent positives confined to section edges, folds, or necrotic regions, and compare them with a no primary control to identify staining artefacts (standard IHC practice). With HRP chromogenic detection, a peroxidase block helps assess endogenous peroxidase background; GUSB itself is annotated as a glycosidase, not a peroxidase (caption A02234; UniProt P08236 function; standard IHC practice).
Boster reagents
Best GUSB / Beta-glucuronidase IHC Antibodies
Two anti-GUSB antibodies have human paraffin-section IHC images (catalog IHC captions); A02234-1 also has an A431-cell IF image (catalog IF caption). A02234 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 has IHC images from formaldehyde-fixed, paraffin-embedded human liver and colon carcinoma, using EDTA pH 9 retrieval and 1:100 primary antibody (A02234 IHC captions). A02234-1 has an IHC image from paraffin-embedded human liver cancer using EDTA pH 8 retrieval and 2 μg/ml primary antibody, plus an IF image from A431 cells (A02234-1 IHC and IF captions).
Which to pick: For tissue IHC, choose A02234 to match its formaldehyde-fixed human liver or colon carcinoma examples (A02234 IHC captions), or A02234-1 to match its human liver cancer example; the latter caption does not report a fixative (A02234-1 IHC caption). For IF/ICC, choose A02234-1 because both applications are listed and its A431-cell IF image uses 2 μg/ml (A02234-1 applications; IF caption). For mouse samples, A02234 is the listed human/mouse-reactive polyclonal option with IHC-P listed, though its IHC images show human tissue (A02234 reactivity; dilution listing; applications; 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-fixed sections are documented (selected-SKU IHC image A02234); keep fixation consistent (standard IHC practice; not target-specific); Caveat: Endogenous peroxidase can mimic HRP/DAB staining (standard IHC practice); Isoform / epitope: 3 isoforms; mature chain begins at residue 23 (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.