CDH17 / Cadherin-17 · IHC design guide

Design Immunohistochemistry for CDH17

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).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for CDH17 (IHC for CDH17): expected localisation Membranous and cytoplasmic in GI glands (HPA tissue IHC), antibody A06389-1, validated IHC image, and IHC protocol steps
Printable CDH17 IHC protocol sheet — expected localisation Membranous and cytoplasmic in GI glands (HPA tissue IHC), antibody A06389-1, controls and protocol steps. Open the full CDH17 IHC guide →

CDH17 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended CDH17 IHC & IF Protocols

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).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human rectal cancer tissue; fixative not specified (datasheet A06389-1)
FixationImage fixative and duration unreported (datasheet A06389-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A06389-1); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A06389-1)
Primary antibodyRabbit anti-CDH17, 1-2 μg/ml (datasheet A06389-1)
Primary incubationOvernight at 4 °C (datasheet A06389-1)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A06389-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultCDH17-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.
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A06389-1); the published protocols used different retrieval conditions (PMC12294824; PMC6970965; PMC4360917).
Section 2

What Is the Expected CDH17 Staining Pattern?

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).

What am I looking at on my slide?
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.
💡Expected CDH17 appearanceCall a result convincing when intestinal endocrine cells show strong, distinct membrane-associated staining with permissible cytoplasmic staining (HPA: high in specified intestinal endocrine cells; membranous and cytoplasmic gland pattern; UniProt Q12864 topology); diffuse section-wide or nuclear-only color is a suspect pattern, not a CDH17-positive call (HPA: tissue IHC; general IHC practice).
How each factor affects the staining
Compartment and epitope contextCDH17 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 selectionIntestinal 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 stainingHPA 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 validationFour 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.
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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.

Sample controls for CDH17 IHC & IF

🧪Run rectum first and require staining in its endocrine cells (HPA: rectum endocrine cells High). Use adrenal gland as the negative tissue because its glandular cells are not detected, and expect adjacent stromal cells on the rectum slide to show only background, without crisp membrane staining (HPA: adrenal gland glandular cells Not detected; HPA: rectum endocrine cells High; UniProt Q12864: cell membrane).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CDH17 in A-549, CACO-2, with annotated localisation: Cell Junctions (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, concentration-matched rabbit IgG isotype control, and CDH17 knockout material or validated immunizing-peptide competition as a biological negative (selected IHC caption: rabbit primary; standard IHC controls). For the biotinylated secondary and SABC-DAB detection scheme, quench endogenous peroxidase and check for endogenous biotin in rectum (selected IHC caption: biotinylated secondary and SABC-DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06389-1 paraffin-section caption does not state a fixative (selected IHC caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used, but whether retrieval is required is unreported; the supplied evidence also does not establish whether frozen sections or IF are easier (selected IHC caption: EDTA retrieval; HPA subcellular: ICC-IF images in A-549 and CACO-2). For rectal IF, assess tissue autofluorescence before interpreting junctional signal (standard IF practice; HPA subcellular: Cell Junctions supported).

HPA tissue IHC evidence for 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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced CDH17 IHC Tips

Troubleshoot CDH17 staining in paraffin sections by checking retrieval, compartment, tissue controls and detection before interpreting signal intensity.

Which retrieval condition should I start with for CDH17 IHC?
Start with heat-mediated retrieval in EDTA pH 8.0 for the catalog antibody (datasheet A06389-1). The selected paraffin-section example used this condition before staining human rectal cancer tissue (caption A06389-1). Keep section thickness, heating and cooling consistent across the test and control slides so differences in signal remain interpretable (standard IHC practice). If staining is weak, first check that retrieval actually reached the tissue and compare with a known positive gastrointestinal section (standard IHC practice; UniProt Q12864 tissue specificity). Record any subsequent retrieval adjustment alongside the antibody concentration and detection method, since changing several steps together obscures the cause (standard IHC practice).
Could fixation explain weak or uneven CDH17 staining?
The selected image documents a paraffin-embedded section but does not state its fixative, so target-specific fixation sensitivity is unknown (caption A06389-1). Record the specimen’s fixative and processing history, then compare affected sections with a similarly processed positive control (standard IHC practice). Inspect whether staining loss tracks folds, damaged tissue or uneven processing before changing antibody conditions (standard IHC practice). Use the documented EDTA pH 8.0 retrieval condition as the starting point for this antibody, while holding detection settings constant (datasheet A06389-1; standard IHC practice). Do not attribute a fixation effect to CDH17 solely from its membrane topology or published tissue pattern (UniProt Q12864 topology; HPA tissue IHC).
Where should convincing CDH17 staining appear in a tissue section?
Expect a prominent membrane-associated pattern because CDH17 is a cell-membrane protein with a transmembrane segment at residues 788–808 (UniProt Q12864). Cell-junction localisation is supported in subcellular imaging, while HPA tissue IHC describes membranous and cytoplasmic staining in gastrointestinal glands (HPA subcellular; HPA tissue IHC). Assess membrane outlines in intact cells and compare cytoplasmic signal with the pattern in a positive control processed in the same run (standard IHC practice; HPA tissue IHC). Nucleoplasmic localisation is listed as uncertain, so isolated nuclear chromogen should prompt a detection and morphology check before interpretation (HPA subcellular; standard IHC practice). Document the compartment separately from overall staining intensity (standard IHC practice).
How should an unknown antibody epitope affect CDH17 troubleshooting?
Do not assume which side of the membrane this catalog antibody recognises: its epitope is not specified in the supplied caption (caption A06389-1). CDH17 has an extracellular region at residues 23–787 and a short cytoplasmic region at 809–832 (UniProt Q12864 topology). It has 8 annotated glycosylation sites and no annotated isoforms, facts relevant when evaluating epitope access or comparing antibodies (UniProt Q12864). If staining differs between antibody preparations, check each reagent’s stated immunogen or epitope before changing retrieval conditions (standard IHC practice). Compare staining on matched sections with the same detection conditions, and avoid assigning the difference to a particular glycosylation site without direct evidence (standard IHC practice; UniProt Q12864).
How can IF help check a questionable CDH17 IHC pattern?
Use IF as a separate localisation check, since the selected catalog evidence describes chromogenic staining of a paraffin section, not an IF protocol (caption A06389-1). Multiplex CDH17 with an independently validated marker for the expected gastrointestinal gland cell population, and assess whether signals occupy the same cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and a channel with low tissue autofluorescence; include unstained and single-stain controls before judging overlap (standard IF practice). CDH17 spans residues 788–808; decide whether to permeabilise only after establishing whether the IF antibody recognises its extracellular or cytoplasmic side (UniProt Q12864 topology; standard IF practice). Do not transfer the IHC retrieval condition to IF without validation (datasheet A06389-1; standard IF practice).
What should I check when CDH17 chromogen appears throughout the section?
Compare the stained slide with a no-primary control to identify signal from secondary detection, endogenous enzyme activity or tissue pigment (standard IHC practice). The selected workflow used 10% goat serum, biotinylated goat anti-rabbit secondary antibody and a streptavidin-biotin complex with DAB (caption A06389-1). Check that the general peroxidase-block step worked, and evaluate background from the biotin-based detection system before increasing blocking or reducing primary antibody (standard IHC practice; caption A06389-1). Keep the documented primary concentration of 2 μg/ml as a reference while changing one variable at a time (caption A06389-1; standard IHC practice). Diffuse staining outside plausible cellular compartments warrants review against tissue morphology and controls (UniProt Q12864; standard IHC practice).
How should I score CDH17 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment before analysis, because supported cell-junction localisation and reported cytoplasmic tissue staining should not be pooled without a stated rule (HPA subcellular; HPA tissue IHC; standard IHC practice). For comparable regions, report the percentage of positive cells and an intensity-based H-score, with membrane and cytoplasmic scores recorded separately (standard IHC practice). Normalise cell counts to the number of eligible cells examined, or positive-cell density to the analysed area in mm² (standard IHC practice). Exclude folds, necrotic areas and section edges using the same criteria for every specimen (standard IHC practice). Include a gastrointestinal positive control in each run and document its staining alongside the specimen scores (UniProt Q12864 tissue specificity; standard IHC practice).
When is apparent CDH17 positivity more likely to be artefact?
Give greatest weight to staining in intact gastrointestinal gland cells with a plausible membrane or junctional pattern, while recording associated cytoplasmic signal separately (HPA tissue IHC; HPA subcellular; standard IHC practice). CDH17 is reported in the gastrointestinal tract and pancreatic duct, whereas UniProt reports no detection in kidney, lung, liver, brain, adrenal gland or skin (UniProt Q12864 tissue specificity). Isolated nuclear signal deserves caution because the reported nucleoplasmic location is uncertain (HPA subcellular). Signal confined to cut edges, necrotic areas or the no-primary control suggests a processing or detection artefact (standard IHC practice). Check endogenous peroxidase and the biotin-based DAB system when brown deposits appear outside intact cells (caption A06389-1; standard IHC practice).
Boster reagents

Best CDH17 / Cadherin-17 IHC Antibodies

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).

Real IHC data IHC analysis of LI Cadherin/CDH17 using anti-LI Cadherin/CDH17 antibody (A06389-1). LI Cadherin/CDH17 was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LI Cadherin/CDH17 Antibody (A06389-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-LI Cadherin/CDH17 Antibody
Cat # A06389-1

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).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry Q12864 (CAD17_HUMAN, Cadherin-17).
  2. Human Protein Atlas. CDH17 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. CDH17 subcellular location (ICC-IF): Mainly localized to the cell junctions. In addition localized to the nucleoplasm..
  4. Human Protein Atlas. CDH17 antibody validation summary (4 antibodies).
  5. Evidence for the Prognostic Value of CDH17 Expression in Colorectal Carcinoma. International journal of molecular sciences 2025 — PMC12294824.
  6. (111)In-labeled anti-cadherin17 antibody D2101 has potential as a noninvasive imaging probe for diagnosing gastric cancer and lymph-node metastasis. Annals of nuclear medicine 2020 — PMC6970965.
  7. Cadherin 17 is a sensitive and specific marker for metanephric adenoma. The American journal of surgical pathology 2015 — PMC4360917.
  8. Cadherin 17 is frequently expressed by 'sclerosing variant' pancreatic neuroendocrine tumour. Histopathology 2015 — PMC4302003.
  9. PubMed PMID:8153632 — UniProt-cited evidence.
  10. PubMed PMID:16421571 — UniProt-cited evidence.
  11. PubMed PMID:15489334 — UniProt-cited evidence.