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Plan KRT7 staining in paraffin sections using the catalog antibody's IHC range of 2–5 μg/ml (datasheet A02416-2). Assess cytoplasmic and membranous signal in glandular epithelium against the reported normal tissue pattern (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic and membranous in glandular epithelium (HPA tissue IHC) | |
| Staining pattern | Glandular epithelial cytoplasmic and membranous staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02416-2) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02416-2) | |
| Caveat | Normal glandular epithelium can stain strongly (HPA tissue IHC) | |
| Regulation | Tissue-dependent epithelial expression (HPA tissue IHC) | |
| Isoform / epitope | No listed isoforms; intracellular epitope access matters (UniProt) |
The catalog antibody protocol uses EDTA retrieval (datasheet A02416-2). The published IHC protocols below provide conditions for urothelial tumours, sarcoma, and cervical carcinoma (PMC2361501; PMC6751227; PMC9131751).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A02416-2) |
| Fixation | Image fixative and duration unreported (datasheet A02416-2); 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 A02416-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02416-2) |
| Primary antibody | Rabbit anti-KRT7, 2-5 μg/ml (datasheet A02416-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02416-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02416-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KRT7-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression mainly in glandular epithelium. No signal in the no-primary control. |
KRT7 is a cytoplasmic intermediate filament protein with no transmembrane segment (UniProt P08729: localisation, topology). In paraffin IHC, expect cytoplasmic and sometimes membranous staining, mainly in glandular epithelium (HPA: tissue IHC profile). HPA rates the tissue pattern Enhanced while reporting medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability); interpret signal by cell type and compartment.
| Strong cytoplasmic staining in breast glandular cells or bronchial respiratory epithelium. | This fits two observed high-expression patterns (HPA: breast glandular cells, High; bronchus respiratory epithelial cells, High). Assess the stained cells within their tissue context, since the HPA observations identify specific cell populations. |
| Cytoplasmic staining with an apparent membranous component in glandular epithelium. | This can fit the reported paraffin IHC profile (HPA: cytoplasmic and membranous expression mainly in glandular epithelium). KRT7 has no transmembrane segment, so a membrane-like edge does not establish membrane insertion (UniProt P08729: topology). |
| Nuclear-only staining, with little or no cytoplasmic signal in expected positive cells. | Treat this as a compartment mismatch requiring investigation: UniProt places KRT7 in the cytoplasm, and HPA supports cytosol and intermediate filaments by ICC-IF (UniProt P08729: localisation; HPA: subcellular). |
| Signal concentrated in adipocytes or bone-marrow hematopoietic cells. | Those cell populations are reported as not detected (HPA: adipocytes, Not detected; bone marrow hematopoietic cells, Not detected). Check for cross-reactivity or endogenous detection activity before scoring such signal as KRT7 (standard IHC practice). |
| Widespread haze across cells and surrounding tissue, without a discernible cell pattern. | Diffuse background cannot establish the reported cell-specific pattern (HPA: tissue IHC profile). Inspect the no-primary control and detection steps for nonspecific signal, then reassess the positive cells (standard IHC practice). |
| Cell compartment and topology (UniProt P08729: localisation, topology). | KRT7 is cytoplasmic and lacks a transmembrane segment (UniProt P08729: localisation, topology). Interpret a membrane-like outline alongside cytoplasmic staining; do not infer a membrane-spanning protein from the IHC appearance alone. |
| Positive tissue and cell choice (HPA: tissue IHC). | HPA reports High staining in breast glandular cells, bronchial respiratory epithelial cells, kidney collecting ducts and lung alveolar type I cells (HPA: listed cell-specific levels). Select a documented cell population when judging a positive control. |
| Negative and low-level comparators (HPA: tissue IHC). | HPA lists adipocytes and bone-marrow hematopoietic cells as Not detected, while stomach glandular cells are Low (HPA: listed cell-specific levels). A weak stomach result therefore has a different interpretation from failure in a documented High population. |
| Antibody evidence (HPA: antibody validation; tissue IHC reliability). | HPA007272 and CAB000028 have Enhanced IHC validation; CAB079970 and CAB080115 have Supported IHC validation (HPA: antibody list). HPA also reports medium antibody–RNA consistency for the tissue profile (HPA: reliability description). |
| IF/ICC: where should signal appear? (HPA: subcellular). | Expect cytosol and intermediate-filament localisation in ICC-IF; HPA marks filaments Approved and cytosol Supported (HPA: subcellular). This is an interpretation cue for IF/ICC, whose preparation needs its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a documented High population, such as breast glandular cells (HPA: breast, High). | The staining run or selected cell population may be unsuitable; HPA's level is cell-specific (HPA: tissue IHC). | Confirm that the expected cells are present, then review the positive control, antibody dilution, retrieval and detection run records (standard IHC practice). No KRT7-specific retrieval condition is established by these sources. |
| Weak staining in stomach glandular cells (HPA: stomach glandular cells, Low). | Weak signal may reflect the reported low expression, rather than a failed run (HPA: stomach, Low). | Compare with a documented High population on a concurrently processed control, and score the stomach cells in their own context (HPA: tissue IHC; standard IHC practice). |
| Nuclear-only signal in otherwise expected epithelial cells. | The compartment conflicts with cytoplasmic KRT7 localisation (UniProt P08729: localisation; HPA: subcellular). | Check the no-primary control, counterstain and detection specificity; require a reproducible cytoplasmic component before accepting the result (standard IHC practice). |
| Apparent positivity in adipocytes or bone-marrow hematopoietic cells. | HPA reports these cells as Not detected; unexpected signal could reflect cross-reactivity or endogenous detection activity (HPA: tissue IHC; standard IHC practice). | Compare no-primary and appropriate detection controls, inspect cell identity, and repeat with an independently validated IHC antibody if needed (standard IHC practice; HPA: antibody validation). |
| Diffuse staining obscures epithelial borders and cell identity. | Background from blocking, antibody concentration or detection steps can obscure a cell-specific pattern (standard IHC practice; HPA: tissue IHC profile). | Review blocking and washes, compare the no-primary control, and adjust the staining workflow only after locating the background source (standard IHC practice). |
| Two antibodies give different tissue patterns. | HPA assigns different validation levels to listed IHC antibodies, while its tissue profile has medium antibody–RNA consistency (HPA: antibody list; reliability description). | Compare both stains in the same documented High and Not detected cell populations, then check each antibody's IHC validation and controls before resolving the discrepancy (HPA: tissue IHC; standard IHC practice). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Ciliated epithelial cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot KRT7 staining in paraffin sections by checking retrieval, cell identity, filament localisation and the controls used for chromogenic detection.
Anti-KRT7 antibodies have IHC images from human breast cancer, prostate and uterus, and mouse and rat liver (catalog IHC captions); IF images show human cancer sections and HeLa cells (catalog IF captions).
A02416-2 shows IHC in human breast cancer and IF in human cancer sections (A02416-2 image captions); M02416-4 shows IHC in human prostate and mouse and rat liver (M02416-4 IHC captions). M02416-1 shows IHC in human breast carcinoma and IF in HeLa cells (M02416-1 image captions); M02416-2 shows IHC in human uterus and IF in HeLa cells (M02416-2 image captions).
Which to pick: For tissue IHC, A02416-2 provides a documented starting point in paraffin human breast cancer sections: EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C; the fixative is unreported (A02416-2 IHC caption). For human IF/ICC, choose rabbit monoclonal M02416-1: IF is pictured in HeLa cells, and IF and ICC are listed applications (M02416-1 IF caption; catalog applications and clonality). For IHC across species, choose rabbit monoclonal M02416-4, with images from human prostate and mouse and rat liver (M02416-4 IHC captions; catalog clonality); its tissue processing and fixatives are unreported (M02416-4 IHC captions).