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- Table of Contents
Plan CDH17 paraffin IHC around the membranous and cytoplasmic staining seen in gastrointestinal glands (HPA tissue IHC). This guide covers the catalog antibody’s IHC conditions (datasheet A06389-1) and epitope placement across the extracellular and cytoplasmic regions (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Membranous and cytoplasmic in GI glands (HPA tissue IHC) | |
| Staining pattern | GI endocrine cells show membranous and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06389-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across paraffin 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 A06389-1) | |
| Caveat | Cytoplasmic signal can complicate membrane-only scoring (HPA tissue IHC) | |
| Regulation | Intestine-enriched expression (HPA tissue IHC) | |
| Isoform / epitope | No isoforms annotated; map extracellular vs cytoplasmic epitopes (UniProt) |
The catalog antibody’s IHC-P protocol is followed by three published CDH17 IHC protocols for colorectal carcinoma, gastric carcinoma, and renal tumor sections (PMC12294824; PMC6970965; PMC4360917).
| Sample | Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A06389-1) |
| Fixation | Image fixative and duration unreported (datasheet A06389-1); 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 A06389-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06389-1) |
| Primary antibody | Rabbit anti-CDH17, 1-2 μg/ml (datasheet A06389-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06389-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06389-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDH17-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Distinct membranous and cytoplasmic expression in gastrointestinal glands. No signal in the no-primary control. |
CDH17 is a cell-membrane protein with one transmembrane segment and a large extracellular region (UniProt Q12864 topology). In paraffin-section IHC, expect membranous and cytoplasmic staining in gastrointestinal glands (HPA: Enhanced tissue IHC; high consistency with RNA expression). HPA reports high staining in endocrine cells of the appendix, colon, duodenum, rectum and small intestine, and medium staining in stomach glandular cells (HPA: tissue IHC).
| Membranous staining, with some cytoplasmic staining, in gastrointestinal glands. | This matches the reported tissue pattern (HPA: Enhanced tissue IHC). Judge the stained cells and compartment together; cytoplasmic staining alone is less persuasive than a pattern that also includes the membrane expected from CDH17 topology (UniProt Q12864 topology). |
| Predominantly nuclear staining, with little or no membrane signal, in a paraffin section. | Treat this as an IHC pattern requiring review: CDH17 is annotated at the cell membrane (UniProt Q12864), whereas the HPA tissue pattern is membranous and cytoplasmic (HPA: tissue IHC). HPA reports uncertain additional nucleoplasmic localization in ICC-IF; that observation does not validate nuclear-only tissue IHC (HPA: subcellular ICC-IF). |
| Strong staining in cells outside the expected gastrointestinal gland pattern. | Check cell identity and the detection controls before calling it CDH17. HPA reports high staining in specified intestinal endocrine cells and medium staining in stomach glandular cells (HPA: tissue IHC); an unexpected cell pattern could reflect cross-reactivity or endogenous detection activity (general IHC practice). |
| Diffuse color across many cells or the surrounding section, obscuring cell borders. | This does not reproduce the distinct glandular membrane and cytoplasmic pattern (HPA: tissue IHC). Background can arise from nonspecific reagent binding, inadequate washing or endogenous detection activity (general IHC practice); compare a no-primary control and inspect the positive tissue pattern. |
| No signal in a known-positive intestinal section. | First confirm that the chosen section contains the relevant endocrine cells: HPA reports high staining in those cells in the appendix, colon, duodenum, rectum and small intestine (HPA: tissue IHC). Then review antibody dilution, retrieval, detection and control performance (general IHC practice); absence of signal alone does not establish absence of CDH17. |
| Compartment and epitope context | CDH17 spans the membrane at residues 788–808, with residues 23–787 extracellular and 809–832 cytoplasmic (UniProt Q12864 topology). The antibody epitope is not supplied, so topology guides localization interpretation but cannot specify antigen retrieval or permeabilisation settings. |
| Tissue and cell selection | Intestinal endocrine cells provide reported high-staining examples; stomach glandular cells are reported at medium level, and gallbladder glandular cells at low level (HPA: tissue IHC). Select controls by the annotated cell type, since a tissue name alone does not identify the cells being scored. |
| Low or absent reference staining | HPA reports no detection in adrenal gland glandular cells, bronchial respiratory epithelial cells and several other listed cells (HPA: tissue IHC). UniProt also reports no detection in kidney, lung, liver, brain, adrenal gland and skin (UniProt Q12864 tissue specificity). Use these as contextual expectations, not proof that every cell in a section must be blank. |
| Antibody validation | Four listed antibodies have Enhanced IHC status: HPA023614, HPA023616, HPA026556 and CAB025143 (HPA: antibody validation). This supports the reported IHC pattern through independent antibodies or orthogonal data; it does not establish that another catalog antibody has the same specificity or optimal conditions. |
| IF/ICC: should nuclear signal be expected? | Cell junctions are the supported main ICC-IF location; additional nucleoplasmic localization is uncertain (HPA: subcellular ICC-IF). Interpret IF/ICC on its own guide page. The ICC-IF observation should not be used as a protocol option or as confirmation of nuclear staining in paraffin-section IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| Intestinal positive control has no stain. | The sampled area may lack the annotated endocrine cells (HPA: high in specified intestinal endocrine cells), or an IHC step may have failed (general IHC practice). | Locate the relevant cells on the section; verify control and detection reagents, antibody dilution and retrieval against the antibody's validated IHC-P instructions (general IHC practice). Do not infer a CDH17-specific retrieval defect from these sources. |
| Stomach control looks weaker than intestine. | HPA reports medium staining in stomach glandular cells and high staining in endocrine cells of the listed intestinal tissues (HPA: tissue IHC). | Score each annotated cell population against its own reference level (HPA: tissue IHC). If a strong control is needed, use a listed high-staining intestinal cell population. |
| Nuclei dominate the chromogenic signal. | Nuclear-only tissue IHC conflicts with the membrane annotation and reported glandular pattern (UniProt Q12864 topology; HPA: tissue IHC). | Check counterstain, detection background and a no-primary control (general IHC practice). Keep the uncertain ICC-IF nucleoplasmic observation separate from the IHC interpretation (HPA: subcellular ICC-IF). |
| Unexpected cells stain strongly. | The cell distribution differs from the reported gastrointestinal gland pattern (HPA: tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm cell identity and compare positive tissue, negative tissue and no-primary controls (general IHC practice). Treat the unexpected pattern as unresolved until the controls support it. |
| Color is diffuse or cell borders are hard to resolve. | Nonspecific background can obscure the distinct membranous and cytoplasmic pattern (HPA: tissue IHC; general IHC practice). | Inspect a no-primary control, blocking and wash performance, and detection-reagent background (general IHC practice). Reassess localization only after the background is controlled. |
| A presumed negative section shows focal color. | Reference absence is cell- and tissue-specific (HPA: tissue IHC; UniProt Q12864 tissue specificity); focal color alone does not identify its source. | Identify the stained cells and compartment, then compare the negative and no-primary controls with a positive gastrointestinal section (general IHC practice). Investigate focal signal before scoring it as CDH17. |
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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | High | Protein (IHC) | HPA → |
| Small intestine | Endocrine cells | High | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot CDH17 staining in paraffin sections by checking retrieval, compartment, tissue controls and detection before interpreting signal intensity.
The IHC-validated antibody has paraffin-section images from human rectal cancer and mouse and rat colon (catalog IHC captions); IF images show CACO-2 cells and human intestinal cancer sections (catalog IF captions).
A06389-1 is the SKU rendered here, with IHC images from paraffin-embedded human rectal cancer and mouse and rat colon (A06389-1 IHC captions). Its IF images show CACO-2 cells and paraffin-embedded human intestinal cancer sections (A06389-1 IF captions).
Which to pick: Choose A06389-1 for tissue IHC: its caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody in paraffin-embedded human rectal cancer (A06389-1 IHC caption); the fixative is unreported (A06389-1 IHC caption). Choose A06389-1 for IF/ICC, supported by images of CACO-2 cells and human intestinal cancer sections (A06389-1 IF captions). It is also the documented cross-species IHC choice, with mouse and rat colon images (A06389-1 IHC captions); M06389 lists human IHC and a monoclonal clone but has no IHC image in the payload (M06389 catalog entry).