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- Table of Contents
Plan DSG1 chromogenic IHC in paraffin sections using the observed membranous pattern in squamous epithelia as a reference (HPA tissue IHC). Compare epidermal layers when scoring, since DSG1 expression is highest in the granular layer (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Squamous epithelial cell membranes (HPA tissue IHC) | |
| Staining pattern | Membranous staining in squamous epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02655-1) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M02655-1) | |
| Caveat | Epidermal staining varies by layer (UniProt) | |
| Regulation | Highest in the epidermal granular layer (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet M02655-1). Four published DSG1 IHC protocols provide tissue-specific methods (PMC12283034; PMC12599537; PMC7436288; PMC9508372).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M02655-1) |
| Fixation | Image fixative and duration unreported (datasheet M02655-1); 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 M02655-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02655-1) |
| Primary antibody | Rabbit monoclonal (clone ED-4) anti-DSG1, 1:50 (datasheet M02655-1) |
| Primary incubation | Overnight at 4 °C (datasheet M02655-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02655-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DSG1-positive staining in squamous epithelial cells of cervix (HPA tissue IHC: High). HPA tissue profile: Distinct membranous expression in squamous epithelia. No signal in the no-primary control. |
In paraffin sections, DSG1 should show distinct cell-border staining in squamous epithelia (HPA: tissue IHC profile). It is a transmembrane desmosomal protein with extracellular and cytoplasmic regions (UniProt Q02413 topology). Expect staining in suprabasal epidermis, strongest toward the granular layer, and in suprabasal esophageal epithelium (UniProt Q02413 tissue specificity). HPA rates its tissue IHC pattern Enhanced, citing consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Distinct cell-border staining in squamous epithelium, with strong skin staining toward the surface (HPA: tissue IHC; UniProt Q02413 tissue specificity). | This fits DSG1 at desmosomal cell junctions (UniProt Q02413 subcellular location). HPA reports high staining in skin corneal-layer cells and in squamous epithelial cells of cervix, oral mucosa, and vagina (HPA: tissue IHC). |
| Predominantly nuclear or diffuse cytoplasmic staining without convincing cell borders (HPA: tissue IHC profile). | Reassess as a possible artefact because the documented tissue IHC pattern is membranous (HPA: tissue IHC profile). UniProt also lists nucleus and cytoplasm, so compartment alone cannot prove that every such signal is false (UniProt Q02413 subcellular location). |
| Strong staining in cells outside the expected squamous epithelial pattern (HPA: tissue IHC profile). | Consider cross-reactivity or endogenous detection activity, then compare a known-positive squamous tissue with a negative control (standard IHC practice). Cell identity matters: HPA reports no detection in adipocytes of adipose tissue or hematopoietic cells of bone marrow (HPA: tissue IHC). |
| Widespread haze or staining that obscures cell borders (HPA: tissue IHC profile). | Treat the slide as difficult to interpret until background is reduced (standard IHC practice). A broad deposit does not reproduce the distinct membranous pattern reported by HPA, even if some squamous cells appear positive (HPA: tissue IHC profile). |
| No epithelial cell-border signal in a known-positive squamous tissue (HPA: tissue IHC). | Check tissue preservation, retrieval, antibody and detection controls before calling DSG1 absent (standard IHC practice). Skin, oral mucosa, cervix, and vagina have high reported staining; esophageal squamous cells have medium staining (HPA: tissue IHC). |
| Tissue and epithelial layer (HPA: tissue IHC; UniProt Q02413 tissue specificity). | Select and score the relevant squamous cells: HPA reports high staining in skin corneal-layer cells and several squamous epithelia, while UniProt places epidermal expression throughout suprabasal layers, highest in the granular layer (HPA: tissue IHC; UniProt Q02413 tissue specificity). |
| Antibody validation (HPA: antibody IHC validation). | HPA022128 and HPA074188 are rabbit polyclonal antibodies with Enhanced IHC status (HPA: antibody IHC validation). That supports the reported tissue pattern; it does not establish identical performance for another antibody or an unreported epitope (HPA: antibody IHC validation). |
| Epitope location (UniProt Q02413 topology). | DSG1 spans the membrane at residues 549–569, with extracellular residues 50–548 and cytoplasmic residues 570–1049 (UniProt Q02413 topology). The supplied antibody records give no epitope, so they cannot predict which region either antibody recognizes (HPA: antibody records). |
| Processing and isoforms (UniProt Q02413 processing and isoforms). | UniProt lists a signal peptide, propeptide, mature chain beginning at residue 50, and two isoforms (UniProt Q02413 processing and isoforms). These features warrant checking an antibody’s stated immunogen when available; the supplied antibody records do not identify its epitope or isoform coverage (HPA: antibody records). |
| IF/ICC: what pattern is expected? (HPA: subcellular summary). | A membrane pattern is the limited expectation from HPA’s IF subcellular summary and UniProt’s desmosomal location (HPA: subcellular summary; UniProt Q02413 subcellular location). HPA supplies no ICC/IF image cell lines or ICC validation for these antibodies, so this IHC evidence cannot establish an IF/ICC result (HPA: subcellular and antibody records). |
| Situation | Likely cause | Next action |
|---|---|---|
| Skin positive control lacks cell-border staining (HPA: high skin staining). | An IHC workflow or reagent failure is possible; absence of signal alone does not identify the failed step (standard IHC practice). | Inspect tissue morphology, follow the selected antibody’s IHC-P instructions, and verify retrieval and detection controls before interpreting the sample (standard IHC practice). |
| Esophagus is weaker than skin (HPA: tissue IHC). | This may match the reported levels: esophageal squamous cells are medium, whereas skin corneal-layer cells are high (HPA: tissue IHC). | Score the same cell type and layer across sections, using the reported tissue levels as context rather than forcing equal intensity (HPA: tissue IHC; standard IHC practice). |
| Nuclear staining dominates the squamous epithelium (HPA: tissue IHC profile). | It does not match HPA’s distinct membranous IHC pattern, although UniProt includes nucleus among listed locations (HPA: tissue IHC; UniProt Q02413 subcellular location). | Check whether cell borders also stain, then compare negative controls and an independent IHC-validated antibody before assigning the nuclear signal to DSG1 (standard IHC practice; HPA: antibody IHC validation). |
| Squamous cells and unrelated cells stain similarly (HPA: tissue IHC profile). | Cross-reactivity or endogenous detection activity may explain a pattern that lacks the expected cell-type distinction (standard IHC practice; HPA: tissue IHC). | Review negative controls and assess whether the squamous cell-border pattern persists; investigate detection-system background before scoring unrelated cells as positive (standard IHC practice). |
| Diffuse chromogen obscures epithelial borders (HPA: tissue IHC profile). | Background from blocking, antibody concentration, washing, or detection can make localisation unreliable (standard IHC practice). | Optimize those general IHC steps with appropriate controls and repeat the slide; require resolvable cell borders for the reported pattern (standard IHC practice; HPA: tissue IHC profile). |
| A negative-control tissue shows strong staining (HPA: tissue IHC). | For example, HPA reports no detection in adipocytes of adipose tissue and hematopoietic cells of bone marrow; strong staining there conflicts with that cell-level record (HPA: tissue IHC). | Confirm the stained cell type, examine reagent and detection controls, and withhold a DSG1-specific call until the unexpected signal is resolved (standard IHC practice; HPA: tissue IHC). |
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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Skin | Cells in corneal layer | High | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Troubleshoot DSG1 staining in paraffin sections by checking retrieval, membrane localisation, tissue context and controls before interpreting chromogenic signal.
Two anti-DSG1 antibodies have IHC images from paraffin sections of human skin, human lung cancer tissue, and mouse testis (catalog image captions). Both list IF and human, mouse, and rat reactivity (catalog applications and reactivity).
M02655-1 has IHC images from paraffin sections of human lung cancer tissue and mouse testis (M02655-1 image captions). A02655-1 has an IHC image from paraffin-embedded human skin (A02655-1 image caption); both list IF, but neither provides an IF image (catalog applications and image records).
Which to pick: For tissue IHC, choose M02655-1 for its rabbit monoclonal format and documented paraffin-section staining at 1:50 with EDTA retrieval (M02655-1 catalog entry and image caption); choose A02655-1 for the pictured paraffin-embedded human skin example at 1:200 (A02655-1 image caption). For IF/ICC, M02655-1 lists both applications, while A02655-1 lists IF only; both list 1:50 as the IF dilution (catalog applications and dilutions). Both list human, mouse, and rat reactivity (catalog reactivity); the captions establish paraffin processing for each SKU, but do not report the fixative (catalog image captions).