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HPSE2 / Inactive heparanase-2 · IHC design guide
Design Immunohistochemistry for HPSE2
Plan HPSE2 IHC in paraffin sections using the reported mainly cytoplasmic tissue pattern and high staining in small-intestinal glandular cells (HPA tissue IHC). Interpret the pattern cautiously because at least one variant is secreted (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HPSE2 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 | Tissue: mainly cytoplasmic (HPA tissue IHC); protein: secreted to extracellular matrix (UniProt) | |
| Staining pattern | Mainly cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | A secreted variant may complicate tissue staining patterns (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 4 isoforms; mature chain begins at residue 42 (UniProt) |
Section 1
Recommended HPSE2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published HPSE2 staining conditions for thyroid tissue (PMC4619411), colorectal adenomas (PMC4800254), and breast cancer samples (PMC7054484).
| Sample | Paraffin-embedded human prostate tissue; fixative not specified (datasheet A07511) |
| Fixation | Image fixative and duration unreported (datasheet A07511); 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-HPSE2, 1:50-1:200 (datasheet A07511) |
| 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 | HPSE2-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval rule); the breast study does not specify retrieval buffer or timing (PMC7054484).
Section 2
What Is the Expected HPSE2 Staining Pattern?
HPSE2 is secreted and associated with extracellular space and matrix; it has no transmembrane segment (UniProt Q8WWQ2). In tissue IHC, HPA reports mainly cytoplasmic staining, including High staining in duodenal and small intestinal glandular cells, hepatocytes, and placental trophoblastic cells (HPA tissue IHC). Treat the pattern as provisional: HPA rates its tissue IHC reliability Uncertain and flags possible differences between RNA and protein location (HPA tissue IHC).
| Cytoplasmic chromogen in duodenal or small intestinal glandular cells, hepatocytes, or placental trophoblastic cells. | This matches the cell types and High levels reported by HPA (HPA tissue IHC). Score the named cells and their staining intensity rather than assigning the same score to the whole section. HPA's Uncertain reliability means agreement with this pattern supports interpretation but does not independently establish antibody specificity (HPA tissue IHC). |
| Extracellular staining near otherwise plausible positive cells, with or without cytoplasmic staining. | Extracellular signal is biologically plausible because HPSE2 is secreted and associated with extracellular space and matrix (UniProt Q8WWQ2). It is not the principal tissue pattern described by HPA, which reports mainly cytoplasmic staining (HPA tissue IHC). Assess whether the signal follows tissue structures and persists when the primary antibody is omitted (general IHC practice). |
| Strong, isolated nuclear staining or a continuous cell-surface outline with little expected cytoplasmic signal. | These distributions do not match HPA's mainly cytoplasmic tissue profile or UniProt's secreted, non-transmembrane annotation (HPA tissue IHC; UniProt Q8WWQ2). Treat them as suspect until a control or independent antibody supports them. A membrane outline alone should not be taken as evidence that HPSE2 is a transmembrane protein (UniProt Q8WWQ2). |
| Strong staining confined to adipocytes or oral squamous epithelial cells, while expected positive cells remain unstained. | HPA reports HPSE2 as Not detected in adipocytes of adipose tissue and breast, and in oral mucosal squamous epithelial cells (HPA tissue IHC). This reversal raises concern for nonspecific staining or detection background; it does not prove cross-reactivity. Check the negative control and the identity of the stained cells before interpreting the result (general IHC practice). |
| Uniform color across tissue compartments, or no signal in a section containing a known positive cell type. | Uniform color without a cell-resolved pattern favors background over a convincing HPSE2 result (general IHC practice). An absent signal in duodenal glandular cells or hepatocytes conflicts with HPA's High observations but is not, by itself, proof of HPSE2 absence (HPA tissue IHC). Review controls and detection conditions before scoring the specimen negative (general IHC practice). |
💡Expected HPSE2 appearanceA convincing chromogenic positive shows predominantly cytoplasmic staining in the expected cells, potentially High in small intestinal glandular cells or hepatocytes (HPA tissue IHC); extracellular signal is plausible from the secreted protein annotation (UniProt Q8WWQ2), whereas isolated nuclear staining or uniform section-wide color is suspect (HPA tissue IHC; general IHC practice).
| Secreted location versus observed tissue pattern | UniProt places HPSE2 in extracellular space and matrix and annotates no transmembrane segment (UniProt Q8WWQ2). HPA nonetheless describes mainly cytoplasmic tissue IHC (HPA tissue IHC). Record both observations when scoring; do not require an exclusively extracellular pattern or interpret a membrane outline as established topology. |
| Tissue and cell selection | HPA reports High staining in duodenal, small intestinal, and stomach glandular cells, hepatocytes, and trophoblastic cells; prostate glandular cells are Low (HPA tissue IHC). Choose a reference section by its annotated cell population, since a low-level population offers a less clear visual comparison (HPA tissue IHC). |
| Antibody validation and transcript uncertainty | The supplied HPA antibody, HPA044603, has an Uncertain IHC status, and HPA notes possible splice or transcript discrepancies and complex RNA–protein correlation for a secreted variant (HPA antibody record; HPA tissue IHC). Interpret discordant staining cautiously; RNA enrichment alone does not predict where chromogenic protein signal must appear (HPA tissue IHC). |
| Processing, glycosylation, and isoforms | UniProt annotates a signal peptide at residues 1–41, a mature chain at 42–592, glycosylation sites at 254 and 392, and 4 isoforms (UniProt Q8WWQ2). These features can matter when assessing antibody epitope coverage, but the supplied evidence does not identify this antibody's epitope or establish an effect on IHC staining. |
| ICC/IF evidence boundary | HPA summarizes the subcellular location as secreted but supplies no main ICC/IF location or cell-line images, and the listed antibody has no ICC validation status (HPA subcellular record; HPA antibody record). Tissue IHC observations cannot establish an expected fluorescent cell-line pattern. Consult the separate IF/ICC guide for application-specific design. |
| Situation | Likely cause | Next action |
|---|---|---|
| No chromogenic signal in duodenal glandular cells or hepatocytes. | These cells are reported High by HPA, so a negative section may reflect assay failure, sampling, or true variation; the supplied evidence cannot distinguish them (HPA tissue IHC). | Confirm the relevant cells are present, then review the positive control, antibody dilution, retrieval conditions, and detection reagents (general IHC practice). Do not infer HPSE2-specific retrieval sensitivity from this result. |
| All tissue structures show similar diffuse color. | A section-wide pattern lacks the cell selectivity described in HPA tissue IHC and can arise from nonspecific reagent or detection background (HPA tissue IHC; general IHC practice). | Compare a section processed without primary antibody; review blocking, wash steps, and chromogen development (general IHC practice). Interpret only cell-resolved signal above the control background. |
| Signal is strongest in an HPA-negative cell population. | HPA lists adipocytes and oral squamous epithelial cells as Not detected; strong signal there may represent cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). | Verify cell identity and compare primary-omission and detection controls. Reassess the pattern with independent antibody evidence if available (general IHC practice). |
| Staining appears only in nuclei or as a sharp cell-surface rim. | That pattern conflicts with HPA's mainly cytoplasmic tissue profile; UniProt annotates HPSE2 as secreted and lacking a transmembrane segment (HPA tissue IHC; UniProt Q8WWQ2). | Check whether the same pattern appears in the negative control and whether expected positive cells stain. Withhold a localization claim unless independent evidence supports it (general IHC practice). |
| RNA enrichment suggests a positive tissue, but IHC is weak. | HPA reports tissue-enhanced RNA in brain, cervix, and vagina, while some annotated cell populations there have Low protein staining; HPA also flags complex RNA–protein correlation (HPA tissue IHC). | Compare protein staining with the HPA annotation for the same cell type, such as Low vaginal squamous epithelial staining, rather than treating tissue-level RNA as a protein control (HPA tissue IHC). |
| Can this IHC pattern set the expected result for ICC/IF? | HPA provides no ICC/IF cell-line images or main subcellular location for HPSE2, and the listed antibody has no ICC validation status (HPA subcellular record; HPA antibody record). | Use the separate IF/ICC guide to design and assess that application. Treat the tissue IHC pattern as context, not as an established fluorescent reference (HPA tissue IHC; HPA subcellular record). |
Experiment evidence
IHC & IF images showing HPSE2 staining
A07511 · IHCImmunohistochemistry of paraffin-embedded human prostate using HPSE2 antibody at dilution of 1:100 (40x lens).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172987-HPSE2/tissue/cerebral+cortex#imid_11825207
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172987-HPSE2/tissue/colon#imid_11825117
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172987-HPSE2/tissue/liver#imid_11825118
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172987-HPSE2/tissue/kidney#imid_11825127
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HPSE2 IHC & IF
🧪Run duodenum first and assess staining in glandular cells (HPA: High in duodenal glandular cells). Use oral mucosa squamous epithelial cells as a negative tissue (HPA: Not detected in oral mucosa squamous epithelial cells); on the duodenum slide, assess background in adjacent non-glandular areas without assuming those areas lack secreted HPSE2 (UniProt Q8WWQ2: secreted, extracellular matrix).
Positive control tissue: Duodenum (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 HPSE2; 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; use a host- and isotype-matched control for a monoclonal primary, or host-matched nonimmune IgG for a polyclonal primary; and confirm specificity with HPSE2 knockout tissue or peptide competition when the immunizing peptide is available (standard IHC practice). For chromogenic duodenum IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected A07511 paraffin-section caption does not report its fixative (A07511 tissue-IHC caption). Antigen retrieval dependence is also unreported, so assess retrieval conditions empirically for paraffin IHC (standard IHC practice). The evidence does not establish whether frozen sections or IF/ICC are easier, and HPA lists no ICC-IF cell-line images (HPA subcellular record); duodenal endogenous peroxidase can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for HPSE2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Caution, Splice and/or transcript discrepancy exists. Pending external verification.). 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HPSE2 IHC Tips
Troubleshoot HPSE2 staining in paraffin sections by checking retrieval, controls, compartment, and cell type before comparing signal intensity (UniProt Q8WWQ2; HPA tissue IHC).
How should I retrieve HPSE2 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). After cooling, compare the same IHC-validated antibody on adjacent sections with matched detection and exposure to developer, so retrieval is the main variable (standard IHC practice). If signal remains weak, test an alternative retrieval condition on adjacent sections as a fallback, while watching for tissue damage and increased background (standard IHC practice). Record staining separately in cells and extracellular areas, because HPSE2 is secreted and associated with extracellular matrix, while HPA reports mainly cytoplasmic tissue staining (UniProt Q8WWQ2; HPA tissue IHC).
Can fixation explain variable HPSE2 staining across paraffin blocks?
Target-specific HPSE2 sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (caption A07511). For a controlled comparison, use sections with documented fixation and processing histories, then keep section thickness, retrieval, antibody incubation, and chromogenic development consistent (standard IHC practice). Compare a known positive area within each staining run and note whether poor signal tracks a processing batch rather than a cell population (standard IHC practice). Do not infer fixation tolerance from the reported tissue staining pattern or from HPSE2 secretion and glycosylation, since neither establishes a fixation response (HPA tissue IHC; UniProt Q8WWQ2).
Where should convincing HPSE2 signal appear in tissue sections?
Assess glandular-cell cytoplasm and nearby extracellular regions separately: HPA describes mainly cytoplasmic staining, whereas UniProt places HPSE2 in the extracellular space and matrix (HPA tissue IHC; UniProt Q8WWQ2). HPSE2 has a signal peptide at residues 1–41 and no transmembrane segment, so a crisp membrane-only pattern needs independent confirmation before assignment to HPSE2 (UniProt Q8WWQ2). Compare the stained compartment with matched negative controls and tissue morphology, including luminal material and stromal boundaries (standard IHC practice). Report the compartment and cell type explicitly, because secreted protein can be detected away from its producing cells (UniProt Q8WWQ2; standard IHC interpretation).
Could isoforms or processing explain discordant HPSE2 staining?
HPSE2 has 4 annotated isoforms, a signal peptide at residues 1–41, and glycosylation sites at residues 254 and 392 (UniProt Q8WWQ2). Check the catalog antibody’s stated immunogen or mapped epitope against each isoform before interpreting a negative section as absence of all HPSE2 forms (standard IHC practice; UniProt Q8WWQ2). The supplied evidence does not map this antibody’s epitope or establish isoform-specific staining (supplied record and caption A07511). If staining patterns disagree across antibodies, compare their epitope coverage and use matched controls and an orthogonal assay before assigning the difference to splicing or glycosylation (standard IHC practice).
How can I adapt HPSE2 assessment to multiplex immunofluorescence?
Treat IF as a separate validation exercise: the supplied HPA subcellular record lists no ICC/IF image-bearing cell lines (HPA subcellular). Multiplex HPSE2 with a marker for the expected cell population, such as a glandular epithelial marker when assessing the reported glandular-cell staining, and inspect both cellular and adjacent extracellular signal (HPA tissue IHC; UniProt Q8WWQ2). Choose fluorophores after imaging an unstained section to identify tissue autofluorescence, then include single-label controls for channel bleed-through (standard IF practice). Match permeabilisation to the epitope’s location: test it for an intracellular epitope, while evaluating an accessible extracellular epitope without it; the supplied evidence does not identify this antibody’s epitope (standard IF practice; supplied record).
What should I change when HPSE2 chromogenic staining looks diffuse?
Run a no-primary control, confirm peroxidase blocking, and compare background in matched sections before changing antibody concentration (standard chromogenic IHC practice). Separate diffuse extracellular staining from uniform reagent background: HPSE2 is secreted and matrix-associated, while HPA reports mainly cytoplasmic staining in tissue IHC (UniProt Q8WWQ2; HPA tissue IHC). If background rises with stronger retrieval or longer DAB development, shorten that step in a controlled comparison and keep the reference section in the same run (standard IHC practice). Check tissue edges, damaged areas, and endogenous pigment microscopically, because those patterns can mimic a positive chromogen signal (standard IHC practice).
How should I score HPSE2 across heterogeneous IHC sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment and cell population before counting, and record cytoplasmic and extracellular staining separately (HPA tissue IHC; UniProt Q8WWQ2). For cell-associated staining, report the percentage of positive cells and an H-score from 0–300 using intensity categories 0–3; keep thresholds fixed across the comparison (standard IHC scoring practice). For extracellular deposits, measure positive area or signal density per mm² of eligible tissue rather than assigning a cell H-score (standard image-analysis practice). Normalise cell counts to the number of eligible cells and extracellular measurements to analysed tissue area, excluding folds, necrosis, and blank regions by a prespecified rule (standard IHC practice).
How do I distinguish genuine HPSE2 staining from artefact?
A plausible result should fit tissue morphology and the assessed compartment: HPSE2 is secreted and matrix-associated, while HPA reports predominantly cytoplasmic tissue staining (UniProt Q8WWQ2; HPA tissue IHC). Use reported glandular-cell positives and an HPA-reported undetected cell population as comparison points, while respecting HPA’s Uncertain reliability designation (HPA tissue IHC). Question isolated nuclear or sharply membrane-only staining, edge-restricted signal, and necrotic staining unless independent controls support them (UniProt Q8WWQ2 topology; standard IHC interpretation). A no-primary control and adequate peroxidase block help identify detection-system signal, including endogenous enzyme activity, before assigning DAB staining to HPSE2 (standard chromogenic IHC practice).
Boster reagents
Best HPSE2 / Inactive heparanase-2 IHC Antibodies
A07511 has a paraffin-section human prostate IHC image (A07511 image caption) and lists human, mouse, and rat reactivity (catalog: reactivity).
Real IHC data
Anti-Inactive heparanase-2 HPSE2 Antibody
Cat # A07511
A07511 will render as an IHC-listed antibody with human, mouse, and rat reactivity (catalog: applications and reactivity). Its image shows paraffin-embedded human prostate stained at 1:100 (A07511 image caption).
Which to pick: Choose A07511 for paraffin-section tissue IHC based on its human prostate image; the fixative is unreported (A07511 image caption). For cross-species IHC, A07511 lists human, mouse, and rat reactivity, although its IHC image shows human tissue only (catalog: reactivity; A07511 image caption). There is no IF/ICC-listed SKU here; A07511 has no IF image or dilution, and its rabbit host is reported while clonality is unreported (catalog: applications, IF images, IF dilution, host, and clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8WWQ2 (HPSE2_HUMAN, Inactive heparanase-2).
- Human Protein Atlas. HPSE2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. HPSE2 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. HPSE2 antibody validation summary (1 antibodies).
- The Profile of Heparanase Expression Distinguishes Differentiated Thyroid Carcinoma from Benign Neoplasms. PloS one 2015 — PMC4619411.
- Immunohistochemical expression of heparanase isoforms and syndecan-1 proteins in colorectal adenomas. European journal of histochemistry : EJH 2016 — PMC4800254.
- Heparanase-2 protects from LPS-mediated endothelial injury by inhibiting TLR4 signalling. Scientific reports 2019 — PMC6753096.
- miR-15b-5p Promotes Growth and Metastasis in Breast Cancer by Targeting HPSE2. Frontiers in oncology 2020 — PMC7054484.
- PubMed PMID:11027606 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:20560209 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HPSE2 (Inactive heparanase-2). Expected localisation Tissue: mainly cytoplasmic (HPA tissue IHC); protein: secreted to extracellular matrix (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07511 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate tissue; fixative not specified (datasheet A07511); 2. Fixation: Image fixative and duration unreported (datasheet A07511); 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-HPSE2, 1:50-1:200 (datasheet A07511); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: A secreted variant may complicate tissue staining patterns (HPA tissue IHC); Isoform / epitope: 4 isoforms; mature chain begins at residue 42 (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q8WWQ2, HPA tissue IHC · Uncertain, HPA ICC-IF · Secreted, Datasheet A07511, PMC4619411, PMC4800254. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HPSE2/immunohistochemistry-ihc-guide.