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- Table of Contents
Start CDH13 paraffin IHC with the catalog antibody at 1–2 μg/mL (datasheet A01986-1). Assess membranous and cytoplasmic staining in glomeruli, muscle and endothelium (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Membranous and cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Membrane/cytoplasm in muscle, glomeruli, cortex and endothelium (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01986-1) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A01986-1); verify before use. | |
| Caveat | Appendicitis staining does not establish staining in appendix glands (HPA tissue IHC; datasheet A01986-1) | |
| Regulation | Blood-vessel-enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | 5 isoforms; mature chain spans residues 139–693, so check epitope placement (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet A01986-1) is accompanied by one published CDH13 IHC protocol (PMC13114730).
| Sample | Paraffin-embedded human appendicitis tissue; fixative not specified (datasheet A01986-1) |
| Fixation | Image fixative and duration unreported (datasheet A01986-1); 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 A01986-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01986-1) |
| Primary antibody | Rabbit anti-CDH13, 1-2μg/ml (datasheet A01986-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01986-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01986-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDH13-positive staining in cells in glomeruli of kidney (HPA tissue IHC: High). HPA tissue profile: Distinct membranous and cytoplasmic expression in muscle tissues, renal glomeruli, cerebral cortex, in endothelial cells and in basal cells of squamous epithelia. No signal in the no-primary control. |
CDH13 should show membranous and cytoplasmic staining in selected cells, including renal glomerular cells, myocytes, cardiomyocytes and basal cells of squamous epithelia (HPA: Enhanced tissue IHC; HPA: tissue profile). A membrane pattern fits its GPI-anchor annotation and lack of a transmembrane segment (UniProt P55290 topology and keywords). The cell type and compartment matter more than uniform staining across a section (HPA: tissue profile).
| Defined membrane-associated staining, with some cytoplasmic staining, in glomerular cells. | This matches the reported compartments and a High signal in kidney glomerular cells (HPA: tissue IHC). Score the glomerular cells, not the whole kidney section (HPA: tissue IHC). |
| Moderate staining in cardiomyocytes, skeletal myocytes, smooth muscle cells or skin basal cells. | These are reported Medium positive cell populations (HPA: tissue IHC). Heart expression is described as high by UniProt, while the HPA IHC level for cardiomyocytes is Medium; keep those measures distinct (UniProt P55290 tissue specificity; HPA: tissue IHC). |
| Predominantly nuclear staining without a convincing membrane or cytoplasmic pattern. | Treat this as a suspect compartment pattern and investigate artefact before calling it CDH13: the reported IHC pattern is membranous and cytoplasmic, and the supported ICC-IF location is plasma membrane (HPA: tissue IHC; HPA: subcellular). |
| Strong staining in adipocytes or adrenal glandular cells. | These populations are reported as Not detected (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous detection activity, then compare a matched negative control (general IHC practice); staining alone does not distinguish those causes. |
| Diffuse color across tissue and background, or no signal in glomerular cells. | Diffuse color prevents a reliable compartment call (general IHC practice). Absent glomerular staining conflicts with the reported High level, but could reflect an assay failure; check controls before inferring biological absence (HPA: tissue IHC; general IHC practice). |
| Compartment and anchor | Use the membrane-associated pattern as the main spatial check; cytoplasmic IHC staining is also reported (HPA: tissue IHC). CDH13 has no transmembrane segment and carries a GPI-anchor annotation (UniProt P55290 topology and keywords). |
| Cell-specific tissue pattern | A section can contain cells with different expected results: skin basal cells are Medium, whereas adipocytes are Not detected in the listed HPA samples (HPA: tissue IHC). Interpret staining by cell population. |
| Processing and isoforms | UniProt lists signal and propeptide processing, a mature chain at residues 139–693, and 5 isoforms (UniProt P55290 processing and isoforms). Without an antibody epitope, these facts cannot predict which forms the IHC reagent detects. |
| Antibody evidence | Both listed antibodies have Enhanced IHC validation, indicating agreement with independent antibody staining or orthogonal data (HPA: HPA001380 and CAB025863 IHC validation). That supports the reported pattern but does not validate every new specimen or run. |
| Situation | Likely cause | Next action |
|---|---|---|
| Glomerular cells have no detectable stain. | The assay may have failed, or this specimen may differ from the reported High kidney pattern (HPA: tissue IHC); the slide alone cannot separate these possibilities. | Review a known-positive control and the run's retrieval, primary-antibody and detection steps (general IHC practice). Avoid assigning a CDH13-negative result until the controls work. |
| Color is widespread and cell borders cannot be resolved. | Nonspecific primary-antibody binding or insufficient washing can obscure localisation (general IHC practice). | Compare the negative control, review blocking and wash conditions, and reassess antibody concentration against its validated instructions (general IHC practice). No target-specific dilution is supplied. |
| Adipocytes or adrenal glandular cells stain strongly. | Both are listed as Not detected in the specified cells (HPA: tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Check a no-primary control and the detection system, then compare localisation with a reported positive cell population (general IHC practice; HPA: tissue IHC). |
| Signal is mainly nuclear. | That distribution does not match the reported membrane and cytoplasmic IHC profile (HPA: tissue IHC). Background or an antibody-dependent off-target signal needs investigation (general IHC practice). | Inspect controls and morphology, and confirm the compartment pattern with an independently validated IHC antibody if available (general IHC practice; HPA: antibody validation). |
| Cardiomyocyte staining appears weaker than expected from 'high heart expression.' | UniProt describes high heart expression, while HPA reports Medium IHC staining in cardiomyocytes (UniProt P55290 tissue specificity; HPA: tissue IHC). | Use the cell-level HPA IHC observation when judging this slide, alongside a positive control from the same run (HPA: tissue IHC; general IHC practice). |
| Q: How should an IF/ICC membrane signal be interpreted? | A: Plasma membrane is the supported ICC-IF location; images are listed for HaCaT and SiHa (HPA: subcellular). This is location evidence, not an IHC-P protocol. | Assess whether IF signal outlines the plasma membrane and consult the separate IF/ICC guide for workflow choices (HPA: subcellular; general IF practice). |
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 |
|---|---|---|---|---|
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Medium | Protein (IHC) | HPA → |
| Skin | Cells in basal layer | Medium | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle 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 CDH13 staining in paraffin section IHC by checking retrieval, cell identity and membrane localisation before comparing staining intensity.
Anti-CDH13 antibodies have paraffin-section IHC images from human and mouse tissues (catalog IHC captions) and IF images from SIHA, K562 and 3T3 cells (catalog IF captions). Rat reactivity is listed (catalog reactivity).
A01986-1 has human appendicitis, gastric cancer, renal carcinoma and melanoma paraffin-section IHC images plus SIHA IF images (A01986-1 image captions); M01986-1 has human cervical and squamous cell carcinoma and mouse skeletal muscle paraffin-section IHC images (M01986-1 IHC captions). A10606 has mouse brain IHC and K562 and 3T3 IF images (A10606 image captions).
Which to pick: For human paraffin-section IHC, choose A01986-1 if its documented EDTA pH 8 retrieval and DAB workflow suits the experiment; the fixative is unreported (A01986-1 IHC captions). For IF/ICC, A01986-1 has a SIHA cell image and listed IF/ICC applications (A01986-1 IF caption; catalog applications); A10606 has K562 and 3T3 IF images (A10606 IF captions). For tissue IHC across species, M01986-1 is a rabbit monoclonal, clone 20C70, with listed human, mouse and rat reactivity and human and mouse paraffin-section images; its fixative is unreported (M01986-1 catalog; M01986-1 IHC captions).