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SGSH / N-sulphoglucosamine sulphohydrolase · IHC design guide
Design Immunohistochemistry for SGSH
Plan SGSH staining in paraffin sections using lung macrophages or kidney collecting ducts as high-staining references (HPA tissue IHC). Evaluate granular cytoplasmic staining by cell type, with adipocytes as a documented unstained comparison (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SGSH 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 | Granular cytoplasm (HPA tissue IHC); lysosomal protein (UniProt) | |
| Staining pattern | Granular cytoplasmic staining in several cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04608-1) | |
| 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 | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Lung macrophages stain strongly; score cell types separately (HPA tissue IHC) | |
| Regulation | No expression regulator documented (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain 21–502 (UniProt) |
Section 1
Recommended SGSH IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published SGSH staining methods for mouse paraffin sections and human bronchial biopsies (PMC6881609; PMC6374378).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A04608-1) |
| Fixation | Image fixative and duration unreported (datasheet A04608-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04608-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04608-1) |
| Primary antibody | Rabbit anti-SGSH, 2-5 μg/ml (datasheet A04608-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04608-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04608-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SGSH-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in several cell types. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A04608-1). The bronchial biopsy method used pH 6.0 retrieval (PMC6374378).
Section 2
What Is the Expected SGSH Staining Pattern?
SGSH is a lysosomal enzyme with no transmembrane segment (UniProt P51688 topology). In paraffin-section IHC, expect granular cytoplasmic staining in selected cell types, including kidney collecting ducts, lung macrophages and several glandular epithelia (HPA tissue IHC). HPA rates the tissue staining pattern Enhanced because antibody staining is highly consistent with RNA expression data (HPA tissue IHC reliability).
| Granular cytoplasmic staining is prominent in kidney collecting ducts or lung macrophages. | This fits lysosomal SGSH localisation (UniProt P51688) and the High staining reported for those cells (HPA tissue IHC). Assess the named cells rather than treating every cell in the section as an equivalent positive control (HPA tissue IHC). |
| Staining is predominantly nuclear or outlines the plasma membrane. | That compartment conflicts with the annotated lysosomal location and lack of a transmembrane segment (UniProt P51688 topology). Check the counterstain and compare a no-primary control before assigning the signal to SGSH (general IHC practice). |
| Strong staining appears in adipocytes, skeletal-muscle myocytes or salivary-gland cells. | HPA reports SGSH as Not detected in those respective cell populations (HPA tissue IHC). Review morphology and controls for cross-reactivity or endogenous chromogenic activity (general IHC practice); a single unexpected field does not establish a new SGSH expression pattern. |
| A diffuse deposit covers multiple compartments or obscures cell boundaries. | The expected HPA description is granular cytoplasmic expression, so diffuse staining cannot be scored as that pattern (HPA tissue IHC). Inspect the no-primary control and reagent background, then reassess blocking, washes and detection conditions (general IHC practice). |
| No staining is visible in kidney collecting ducts on a run intended to detect SGSH. | Collecting ducts are reported High by HPA, making them a useful tissue control (HPA tissue IHC). Verify that the ducts are present, then review antibody application, detection and run controls (general IHC practice); absence in one run alone cannot establish biological loss. |
💡Expected SGSH appearanceCall a result positive when the appropriate cells show granular cytoplasmic signal, potentially strong in kidney collecting ducts or lung macrophages (HPA tissue IHC; UniProt P51688); widespread nuclear, membrane-outline or diffuse staining warrants control review (UniProt P51688 topology; general IHC practice).
| Compartment and molecular form | SGSH is annotated in lysosomes, has a signal peptide at residues 1–20 and lacks a transmembrane segment (UniProt P51688 topology and processing). Interpret punctate or granular cytoplasmic staining in that context; those annotations do not predict a specific chromogenic intensity. |
| Tissue and cell selection | HPA reports High staining in adrenal, epididymal, gallbladder and prostate glandular cells, kidney collecting ducts, lung macrophages and placental syncytiotrophoblast cell bodies; breast glandular cells are Medium (HPA tissue IHC). Match the assessment to the cell population, not just the organ. |
| Low and undetected reference populations | Several populations are Low, including cerebral-cortex neurons and rectal glandular cells, while adipocytes and skeletal-muscle myocytes are Not detected (HPA tissue IHC). Low or undetected populations offer different interpretive expectations and should not be treated as interchangeable positive controls. |
| Strength of the IHC evidence | The tissue profile is rated Enhanced for consistency between antibody staining and RNA expression; two listed rabbit polyclonal antibodies, HPA023436 and HPA023451, each have Enhanced IHC status (HPA tissue IHC; HPA antibody validation). This supports the reported pattern without proving every signal in a new run is specific. |
| IF/ICC: is the same pattern independently shown? | HPA provides no main subcellular ICC-IF location or cell-line images, and lists no ICC validation for either antibody (HPA subcellular; HPA antibody validation). Its HUVEC/TERT2 RNA value is an expression measurement, not an IF localisation result (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive kidney section has no signal in collecting ducts. | The expected High collecting-duct signal is missing (HPA tissue IHC); the cause cannot be determined from the negative image alone. | Confirm duct morphology and check a successful run control, antibody application and chromogenic detection steps (general IHC practice). Record the run as inconclusive until the control performs. |
| The whole section shows diffuse brown staining. | A broad deposit does not match the granular cytoplasmic HPA profile (HPA tissue IHC); background from detection reagents or insufficient washing is possible (general IHC practice). | Examine a no-primary control, check endogenous detection activity, and review blocking, washing and chromogen development (general IHC practice). Score only clearly localised cellular signal. |
| Adipocytes or skeletal-muscle myocytes stain strongly. | These populations are reported Not detected (HPA tissue IHC). Cell misidentification or nonspecific signal is possible (general IHC practice). | Verify the stained cell type against morphology and counterstain; compare the no-primary control and a positive cell population on the same run (general IHC practice). |
| Signal is mainly nuclear or traces cell surfaces. | This location conflicts with lysosomal SGSH and its lack of a transmembrane segment (UniProt P51688 topology). | Check focus, counterstain and control staining, then reassess whether granular cytoplasmic signal is present in an HPA-positive population (general IHC practice; HPA tissue IHC). |
| A weak field is being called SGSH-negative. | HPA levels differ by population: breast glandular cells are Medium, while cerebral-cortex neurons are Low (HPA tissue IHC). A weak field alone cannot resolve expression from run performance. | Identify the exact cell population and compare it with a High reference population and run controls before interpreting a negative result (HPA tissue IHC; general IHC practice). |
| An IF/ICC image is being used to validate the IHC call. | The supplied HPA subcellular record has no ICC-IF location or images, and the listed antibodies have no ICC status (HPA subcellular; HPA antibody validation). | Base this paraffin-section call on morphology, IHC controls and the HPA tissue pattern (HPA tissue IHC; general IHC practice). Treat IF/ICC localisation as unverified by the supplied record. |
Experiment evidence
IHC & IF images showing SGSH staining
A04608-1 · IHCIHC analysis of HSS/SGSH using anti-HSS/SGSH antibody (A04608-1). HSS/SGSH was detected in a paraffin-embedded section of human appendiceal
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000181523-SGSH/tissue/cerebral+cortex#imid_5957392
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000181523-SGSH/tissue/adrenal+gland#imid_5957352
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000181523-SGSH/tissue/colon#imid_5957318
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000181523-SGSH/tissue/liver#imid_5957319
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SGSH IHC & IF
🧪Run kidney first and look for staining in collecting ducts (HPA: High in kidney collecting ducts). Use adipose tissue as a negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the kidney slide, treat cells outside collecting ducts that lack staining as internal background references, without assuming every other cell type is SGSH-negative (HPA: High in collecting ducts).
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 carries no ICC-IF cell line for SGSH; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a matched nonimmune rabbit IgG isotype control, and SGSH knockout tissue or a peptide-block control if available (selected-SKU caption: rabbit primary; standard IHC practice). For kidney HRP/DAB staining, quench endogenous peroxidase and assess background before scoring; check tissue autofluorescence if evaluating IF (selected-SKU caption: HRP/DAB; standard IHC/IF practice).
⚠️Feasibility: Paraffin-section IHC is supported by the selected-SKU caption, which used heat retrieval in EDTA at pH 8.0; its fixative is unreported, and no target-specific fixation window or fixation effect is reported in the supplied evidence (selected-SKU caption). Whether frozen sections or IF are easier is unreported, and HPA lists no ICC-IF images for SGSH (HPA subcellular). Kidney autofluorescence can complicate IF interpretation, so assess an unstained section before judging fluorescent signal (standard IF practice).
HPA tissue IHC evidence for SGSH
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Collecting ducts | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SGSH IHC Tips
Troubleshoot SGSH staining by checking retrieval, cellular pattern, controls, and cell-specific scoring in paraffin-section IHC.
What should I change if SGSH staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04608-1). The selected tissue image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (caption A04608-1). Check a concurrently processed positive tissue before changing retrieval conditions; kidney collecting ducts show high staining in the tissue survey (HPA: kidney collecting ducts, High). If staining remains weak, compare retrieval heating and cooling across sections, since uneven heating can alter epitope exposure (standard IHC practice). Record the retrieval conditions and compare granular cytoplasmic signal with background in the same run (HPA: granular cytoplasmic profile).
How should I troubleshoot weak SGSH staining linked to fixation?
The selected image identifies a paraffin-embedded section but does not report its fixative (caption A04608-1). Target-specific fixation sensitivity is therefore unknown; do not attribute weak SGSH staining to a particular fixative or exposure time on this evidence. Compare sections with documented fixation histories using the same EDTA pH 8.0 retrieval and antibody incubation (datasheet A04608-1; standard IHC practice). Include a positive tissue section in each run, such as kidney collecting ducts (HPA: High), to distinguish a run-wide failure from a specimen issue. Examine morphology and staining together, since poorly preserved tissue can make cytoplasmic granules difficult to assess (standard IHC practice; HPA: granular cytoplasmic profile).
What SGSH staining pattern should I expect in paraffin sections?
Look for granular cytoplasmic staining when assessing SGSH in paraffin sections (HPA: tissue IHC profile). This pattern is consistent with its lysosomal location and soluble topology without a transmembrane segment (UniProt P51688: lysosome, topology). Examine the relevant cells within each tissue: kidney collecting ducts and lung macrophages are reported as High (HPA: kidney; HPA: lung). Strong, uniform nuclear or crisp plasma-membrane staining should trigger a review of specificity and detection controls rather than automatic positive scoring (UniProt P51688: lysosome; standard IHC practice). Use a counterstain to identify cells, then judge granular staining within their cytoplasm separately from extracellular deposits (standard IHC practice).
Could SGSH processing or glycosylation change which cells stain?
SGSH has no annotated isoforms, so an apparent change in staining cannot be assigned to an annotated splice isoform (UniProt P51688: isoforms). Its signal peptide spans residues 1–20, while the annotated mature chain spans 21–502 (UniProt P51688: processing). Five glycosylation sites and a modified residue at position 70 are annotated, but the antibody epitope is not specified here (UniProt P51688: glycosylation, modified residues; caption A04608-1). Do not infer which modification affects binding without epitope mapping or a direct comparison (standard IHC practice). If staining varies between sections, first compare retrieval, preservation, and the expected granular cytoplasmic pattern (datasheet A04608-1; HPA: tissue IHC profile).
How can I compare an SGSH IF result with this IHC guide?
Treat IF as a separate assay while using granular cytoplasmic staining as the tissue IHC reference pattern (HPA: tissue IHC profile). For multiplexing, pair SGSH with a marker identifying macrophages in lung or collecting-duct cells in kidney, using validated markers for the chosen specimen (HPA: lung macrophages, High; HPA: kidney collecting ducts, High; standard IF practice). Choose a fluorophore away from the specimen's strongest autofluorescence and assess single-label controls before interpreting overlap (standard IF practice). SGSH is lysosomal and has no transmembrane segment; permeabilisation should permit access to an intralysosomal epitope, although this antibody's epitope is unspecified (UniProt P51688: location, topology; standard IF practice). Do not transfer the paraffin-image retrieval or fixation assumptions to IF (caption A04608-1).
How can I reduce diffuse brown staining without losing SGSH signal?
Compare background with the expected granular cytoplasmic pattern before changing antibody concentration (HPA: tissue IHC profile). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB development (caption A04608-1). Check a no-primary control, washing, and DAB development time to identify detection background (standard chromogenic IHC practice). Include an endogenous peroxidase block when using HRP detection; treat that block as a general assay step rather than SGSH-specific evidence (standard chromogenic IHC practice; caption A04608-1: HRP/DAB). Review tissue edges and damaged areas separately, since deposited chromogen there can exaggerate apparent positivity (standard IHC practice).
How should I score SGSH staining across different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring, because the tissue survey reports High staining in kidney collecting ducts and lung macrophages (HPA: kidney; HPA: lung). For an H-score, record the percentage of cells at intensity grades 0–3 and sum percentage times grade, yielding 0–300 (standard IHC scoring practice). Alternatively, report the percentage of positive cells or positive-cell density per mm² within a defined region (standard IHC scoring practice). Normalize counts to the relevant cell population or viable tissue area, and use the same threshold across slides (standard IHC scoring practice). Record granular cytoplasmic signal separately from diffuse background and exclude necrotic regions from scoring (HPA: tissue IHC profile; standard IHC practice).
When should I question an apparent SGSH-positive result?
A convincing result shows granular cytoplasmic staining in identifiable cells, consistent with the tissue profile and lysosomal location (HPA: granular cytoplasmic profile; UniProt P51688: lysosome). Check the cell identity against context: lung macrophages stain High, whereas bronchial basal cells are reported Not detected (HPA: lung; HPA: bronchus). Predominantly nuclear or sharply membranous staining warrants specificity controls because SGSH has no transmembrane segment and is lysosomal (UniProt P51688: topology, location; standard IHC practice). Treat staining concentrated at section edges or within necrosis cautiously and inspect a no-primary control (standard IHC practice). With HRP/DAB detection, rule out endogenous enzyme activity before attributing persistent brown signal to SGSH (caption A04608-1: HRP/DAB; standard chromogenic IHC practice).
Boster reagents
Best SGSH / N-sulphoglucosamine sulphohydrolase IHC Antibodies
Anti-SGSH antibodies cover imaged human paraffin-section IHC (A04608-1 image caption) and listed human IF/ICC applications without an IF image (M04608-1 applications; IF image alts).
Real IHC data
Anti-HSS/SGSH Antibody ®
Cat # A04608-1
A04608-1 has an IHC image from a paraffin section of human appendiceal adenocarcinoma; its listed reactivity is human, mouse and rat (A04608-1 image caption; catalog reactivity). M04608-1 lists human IHC and IF/ICC applications, but supplies no IHC or IF image (M04608-1 applications; reactivity; image alts).
Which to pick: For tissue IHC, choose A04608-1 for its documented human paraffin-section staining; the fixative is unreported (A04608-1 image caption). For IF/ICC, choose the rabbit monoclonal M04608-1, which lists those applications for human samples (M04608-1 short description; applications; reactivity). For mouse or rat samples, A04608-1 lists reactivity in both species, though its IHC image documents human tissue only (A04608-1 catalog reactivity; image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51688 (SPHM_HUMAN, N-sulphoglucosamine sulphohydrolase).
- Human Protein Atlas. SGSH tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SGSH subcellular location (ICC-IF): Highest expression in HUVEC/TERT2: 46.9 nTPM.
- Human Protein Atlas. SGSH antibody validation summary (2 antibodies).
- Overcoming Limitations Inherent in Sulfamidase to Improve Mucopolysaccharidosis IIIA Gene Therapy. Molecular therapy : the journal of the American Society of Gene Therapy 2018 — PMC6079371.
- Enhancing the Therapeutic Potential of Sulfamidase for the Treatment of Mucopolysaccharidosis IIIA. Molecular therapy. Methods & clinical development 2019 — PMC6881609.
- AAVrh10 Vector Corrects Disease Pathology in MPS IIIA Mice and Achieves Widespread Distribution of SGSH in Large Animal Brains. Molecular therapy. Methods & clinical development 2020 — PMC6940615.
- Expression, activity and localization of lysosomal sulfatases in Chronic Obstructive Pulmonary Disease. Scientific reports 2019 — PMC6374378.
- PubMed PMID:7493035 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SGSH (N-sulphoglucosamine sulphohydrolase). Expected localisation Granular cytoplasm (HPA tissue IHC); lysosomal protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04608-1). Validated IHC antibody: A04608-1 at 2-5 μg/ml (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; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A04608-1); 2. Fixation: Image fixative and duration unreported (datasheet A04608-1); 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: EDTA pH 8.0 (datasheet A04608-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04608-1); 8. Primary antibody: Rabbit anti-SGSH, 2-5 μg/ml (datasheet A04608-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04608-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Lung macrophages stain strongly; score cell types separately (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain 21–502 (UniProt); Regulation: No expression regulator documented (UniProt). Evidence: UniProt P51688, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in HUVEC/TERT2: 46.9 nTPM, Datasheet A04608-1, PMC6881609, PMC6374378. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SGSH/immunohistochemistry-ihc-guide.