This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
HSPG2 / Basement membrane-specific heparan sulfate proteoglycan core protein · IHC design guide
Design Immunohistochemistry for HSPG2
Plan paraffin IHC for HSPG2 using its observed cytoplasmic and extracellular matrix staining (HPA tissue IHC). Start with the catalog antibody at 0.5–1 μg/mL (datasheet: 0.5–1 μg/mL) and assess staining against appropriate controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSPG2 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 | Extracellular matrix and cytoplasm, including endothelium (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in several tissues; matrix positivity (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Heart muscle | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Secretion can complicate RNA–protein agreement in tissue (HPA tissue IHC) | |
| Regulation | Stimulus-dependent regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms listed; map epitopes against endorepellin/LG3 (UniProt) |
Section 1
Recommended HSPG2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published HSPG2 methods for plaque, brain tissue array, and kidney sections (PMC7508285; PMC2904485; PMC12747916).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PB9277); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-HSPG2, 0.5-1μg/ml (datasheet PB9277) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSPG2-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissue types, including endothelial cells. Positivity in extracellular matrix. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the plaque study used citrate at 100 °C for 20 min (PMC7508285).
Section 2
What Is the Expected HSPG2 Staining Pattern?
HSPG2 is secreted into extracellular matrix and basement membranes and has no transmembrane segment (UniProt P98160). HPA reports extracellular matrix positivity and cytoplasmic staining in several tissues, including endothelial cells; cardiomyocyte staining is medium (HPA tissue IHC: Approved).
| ECM positivity and medium cardiomyocyte staining (HPA tissue IHC). | Matches the reported tissue pattern (HPA tissue IHC; UniProt P98160 localisation). |
| Nuclear-only staining. | Suspect artefact; HSPG2 is secreted to ECM and basement membranes (UniProt P98160 localisation). |
| Adrenal glandular cell staining (HPA: Not detected). | Consider cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). |
| Diffuse staining across the section. | Consider nonspecific binding or detection background (general IHC practice). |
| No cardiomyocyte signal in heart muscle (HPA: Medium). | Check assay performance against this reported positive cell type (HPA tissue IHC). |
💡Expected HSPG2 appearancePositive: ECM/basement membrane signal with medium cardiomyocyte staining; nuclear-only signal is suspect (HPA tissue IHC; UniProt P98160).
| Protein processing (UniProt P98160). | Endorepellin spans 3687–4391 and LG3 spans 4197–4391; check antibody epitope (UniProt P98160). |
| IHC antibody validation (HPA antibodies). | Four listed antibodies are IHC Approved; no ICC status is listed (HPA antibodies). |
| IF/ICC expected pattern? | Secreted protein; HPA provides no ICC-IF images or main location (UniProt P98160; HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No heart cardiomyocyte signal (HPA: Medium). | Low assay sensitivity is possible (general IHC practice). | Check a positive control; titrate primary and optimize retrieval (general IHC practice). |
| Diffuse section background. | Nonspecific binding is possible (general IHC practice). | Titrate primary and review blocking and washes (general IHC practice). |
| Nuclear-only staining. | Off-target staining is possible (UniProt P98160 localisation; general IHC practice). | Run a no-primary control and reassess antibody specificity (general IHC practice). |
| Adrenal glandular staining (HPA: Not detected). | Cross-reactivity or endogenous activity is possible (general IHC practice). | Run a no-primary control; check endogenous enzyme blocking (general IHC practice). |
| Signal in a no-primary control. | Detection reagent binding or endogenous activity is possible (general IHC practice). | Review secondary detection and endogenous enzyme blocking (general IHC practice). |
Experiment evidence
IHC & IF images showing HSPG2 staining
PB9277 · IHCAnti-HSPG2 vendor antibody, PB9277,IHC(P) IHC(P): Human Lung Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142798-HSPG2/tissue/cerebral+cortex#imid_3656472
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142798-HSPG2/tissue/colon#imid_3656398
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142798-HSPG2/tissue/liver#imid_3656415
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000142798-HSPG2/tissue/kidney#imid_3656400
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSPG2 IHC & IF
🧪Run heart muscle first and look for staining associated with cardiomyocytes (HPA: Medium in heart muscle cardiomyocytes). Use adrenal gland as the negative tissue, focusing on glandular cells (HPA: Not detected in adrenal gland glandular cells); on the heart slide, nuclei and cells without a specific extracellular deposit should remain clear (UniProt P98160: extracellular matrix and basement membrane localisation).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA Medium)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for HSPG2; derive a cell-line control from the positive tissue's cell type (Cardiomyocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a nonimmune control matched to the primary antibody’s host species and antibody class, with clonality matched where feasible (standard IHC practice). Use knockout tissue or a validated peptide-block control for biological specificity, and quench endogenous peroxidase before chromogenic detection in heart sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval effect are unreported in the supplied evidence; test antigen retrieval on paired paraffin sections (standard IHC practice). The selected PB9277 tissue-IHC caption does not report a fixative (selected-SKU caption: fixative not stated). The evidence does not establish that frozen sections or IF are easier; assess background carefully in heart muscle, including endogenous peroxidase for chromogenic IHC and autofluorescence if using IF (standard IHC/IF practice).
HPA tissue IHC evidence for HSPG2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HSPG2 IHC Tips
Troubleshoot HSPG2 paraffin section IHC by checking retrieval, matrix localization, and compartment aware controls; use IF to clarify ambiguous spatial patterns (UniProt P98160; HPA tissue IHC).
How should I retrieve HSPG2 in paraffin sections when basement membrane staining is weak?
For paraffin IHC, start with citrate pH 6.0 heat induced retrieval at 95–98 °C for 20 min (page retrieval setting). If staining remains weak, compare a longer citrate exposure on serial sections while holding antibody dilution and detection constant; inspect tissue integrity before adopting the change (standard IHC practice). HSPG2 is a secreted basement membrane proteoglycan, so judge retrieval by continuous matrix staining as well as any cellular signal (UniProt P98160; HPA tissue IHC). Include a no primary control and process comparison sections together, because stronger chromogen alone cannot establish improved epitope detection (standard IHC practice).
Can the available evidence tell me whether fixation is masking HSPG2?
Target specific fixation sensitivity is unknown: the PB9277 tissue IHC caption does not state a fixative or fixation time (PB9277 caption). Record the actual fixative and duration for each specimen, then compare similarly processed serial sections before attributing a weak result to fixation (standard IHC practice). Keep retrieval, antibody incubation, and chromogen development matched across that comparison so differences remain interpretable (standard IHC practice). HSPG2 is found in extracellular matrix and basement membrane, but that localization does not establish how its epitope responds to fixation (UniProt P98160). Assess preserved morphology alongside signal, especially where the expected basement membrane outline appears interrupted (standard IHC practice).
Should HSPG2 staining be extracellular, cytoplasmic, or both?
Prioritize extracellular matrix and basement membrane patterns when assessing paraffin IHC, because HSPG2 is secreted and lacks a transmembrane segment (UniProt P98160). Cytoplasmic expression in several tissue types, including endothelial cells, and extracellular matrix positivity are also reported in tissue IHC (HPA tissue IHC). Examine whether a cellular signal follows recognizable cells while adjacent matrix staining traces anatomical boundaries; compare both patterns with a no primary control (standard IHC practice). Do not interpret a diffuse nuclear pattern as the expected HSPG2 distribution without independent validation, since the supplied localization evidence supports secreted protein and matrix deposition (UniProt P98160; HPA tissue IHC).
How could epitope position change what HSPG2 IHC appears to detect?
Ask which HSPG2 region the antibody recognizes before equating every stained structure with intact core protein (standard IHC practice). The record lists the main chain at residues 22–4391, endorepellin at 3687–4391, and LG3 at 4197–4391 (UniProt P98160 processing). An antibody against the C terminal region could detect those processed products as well as the full chain if its epitope is retained; the PB9277 caption supplies no epitope position (UniProt P98160 processing; PB9277 caption). HSPG2 also has 18 listed glycosylation sites and no listed isoforms, so document epitope mapping and avoid assigning distinct staining patterns to an unsupported isoform (UniProt P98160).
How can IF clarify an ambiguous HSPG2 chromogenic staining pattern?
Use IF as a spatial check on an ambiguous paraffin IHC pattern, with a marker for endothelial cells where that cell population is being evaluated (HPA tissue IHC; standard IF practice). Choose a red or far red fluorophore after checking tissue autofluorescence, and acquire single stain controls before interpreting overlap (standard IF practice). HSPG2 is secreted and has no transmembrane segment, so assess matrix accessible staining without routine detergent permeabilization first; permeabilize only if a companion intracellular marker requires it, then check whether matrix signal is retained (UniProt P98160; standard IF practice). The supplied HPA subcellular record has no ICC/IF images, so treat any IF pattern as an assay result requiring its own controls (HPA subcellular).
What controls help distinguish HSPG2 signal from chromogenic background?
Run a no primary control through the same secondary reagent and chromogen development to reveal detection background (standard IHC practice). Include a peroxidase block when using an enzyme based DAB workflow, and compare tissue edges with well preserved interior regions before calling peripheral staining positive (standard IHC practice). Because HSPG2 is an extracellular matrix protein, evaluate whether diffuse deposit follows basement membrane anatomy instead of simply covering damaged or necrotic areas (UniProt P98160; standard IHC practice). If background persists, titrate the primary antibody and shorten chromogen development separately, retaining a section with the same processing conditions as a comparator (standard IHC practice).
How should I score HSPG2 when matrix and cellular staining coexist? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine separate regions for basement membrane or extracellular matrix and for cellular staining, since both matrix positivity and cytoplasmic expression are reported (HPA tissue IHC). For matrix signal, measure positive area or stained length per mm² of evaluable tissue and normalize to the corresponding matrix area or basement membrane length (standard image analysis practice). For a specified cell population, report percent positive cells and, if intensity categories are reproducible, an H score on the same annotated population (standard IHC scoring practice). Apply identical thresholds and chromogen settings across slides, exclude folded or necrotic regions, and record the compartment used for every comparison (standard IHC practice).
When should I distrust an apparent HSPG2 positive result?
Question a predominantly nuclear signal because HSPG2 is reported as secreted and present in extracellular matrix and basement membrane (UniProt P98160; HPA tissue IHC). Check whether staining follows recognizable matrix boundaries or the specific cell population being scored; HPA reports cytoplasmic expression, including endothelial cells, but its listed cell findings vary by tissue (HPA tissue IHC). Strong signal confined to section edges, folds, or necrotic material warrants review against intact regions and a no primary control (standard IHC practice). If staining survives omission of primary antibody in an enzyme based assay, investigate residual endogenous peroxidase or detection reagent background before assigning biological meaning (standard IHC practice).
Boster reagents
Best HSPG2 / Basement membrane-specific heparan sulfate proteoglycan core protein IHC Antibodies
PB9277 has real IHC data from human lung cancer tissue in paraffin sections (catalog: PB9277 figure tag and IHC image caption). No IF data are supplied (catalog: PB9277 IF images).
Real IHC data
Anti-Heparan Sulfate Proteoglycan 2/HSPG2 Antibody ®
Cat # PB9277
PB9277 is listed for human IHC, with an IHC(P) image from human lung cancer tissue (catalog: PB9277 applications, reactivity, and IHC image caption). Its listed IHC dilution is 0.5–1 μg/ml (catalog: PB9277 IHC dilution).
Which to pick: Choose PB9277 for human tissue IHC: it is a rabbit antibody listed for IHC, and its image documents IHC(P) on human lung cancer tissue (catalog: PB9277 host, applications, reactivity, and IHC image caption). The caption supports paraffin sections; the fixative and clonality are unreported (catalog: PB9277 IHC image caption and clone). No IF/ICC application or nonhuman reactivity is listed, so the payload offers no validated IF/ICC or cross-species choice (catalog: PB9277 applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P98160 (PGBM_HUMAN, Basement membrane-specific heparan sulfate proteoglycan core protein).
- Human Protein Atlas. HSPG2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. HSPG2 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. HSPG2 antibody validation summary (4 antibodies).
- Parallel Murine and Human Plaque Proteomics Reveals Pathways of Plaque Rupture. Circulation research 2020 — PMC7508285.
- Single Cell RNA Sequencing Identifies HSPG2 and APLNR as Markers of Endothelial Cell Injury in Systemic Sclerosis Skin. Frontiers in immunology 2018 — PMC6174292.
- Vascular gene expression patterns are conserved in primary and metastatic brain tumors. Journal of neuro-oncology 2010 — PMC2904485.
- Single-cell analysis reveals shared adaptive responses across different types of podocyte injury. Frontiers in immunology 2025 — PMC12747916.
- PubMed PMID:1569102 — UniProt-cited evidence.
- PubMed PMID:1730768 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSPG2 (Basement membrane-specific heparan sulfate proteoglycan core protein). Expected localisation Extracellular matrix and cytoplasm, including endothelium (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PB9277 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Adrenal gland. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PB9277); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-HSPG2, 0.5-1μg/ml (datasheet PB9277); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Secretion can complicate RNA–protein agreement in tissue (HPA tissue IHC); Isoform / epitope: No isoforms listed; map epitopes against endorepellin/LG3 (UniProt); Regulation: Stimulus-dependent regulation not specified (UniProt). Evidence: UniProt P98160, HPA tissue IHC · Approved, HPA ICC-IF · Secreted, Datasheet PB9277, PMC7508285, PMC2904485. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSPG2/immunohistochemistry-ihc-guide.