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- Table of Contents
Plan TP73 paraffin IHC around nuclear staining in subsets of glandular, squamous and respiratory epithelial cells (HPA tissue IHC). Use a documented positive epithelial control and account for DNA damage response and isoform coverage when interpreting signal (HPA tissue IHC; UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Nuclear in tissue; cytoplasmic location also annotated (HPA tissue IHC: nuclear; UniProt: cytoplasm) | |
| Staining pattern | Nuclear in fallopian glandular and squamous/respiratory cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00688) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Nuclear signal may rise after DNA damage (UniProt) | |
| Isoform / epitope | 11 isoforms; check antibody epitope coverage (UniProt) |
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet M00688). Published IHC protocols cover cervical cancer and liver sections (PMC6682548; PMC6920427).
| Sample | Paraffin-embedded human skin cancer tissue; fixative not specified (datasheet M00688) |
| Fixation | Image fixative and duration unreported (datasheet M00688); 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 M00688); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00688) |
| Primary antibody | Rabbit monoclonal (clone EHI-20) anti-TP73, 1:50 (datasheet M00688) |
| Primary incubation | Overnight at 4 °C (datasheet M00688) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00688) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TP73-positive staining in basal cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in a subset of glandular cells in fallopian tube, squamous epithelium and respiratory epithelium. No signal in the no-primary control. |
TP73 is chiefly nuclear in tissue IHC: expect staining in subsets of respiratory basal cells and squamous epithelial cells, with nuclear expression also reported in a subset of fallopian tube glandular cells (HPA tissue IHC). UniProt lists both nucleus and cytoplasm, and reports nuclear accumulation after DNA damage (UniProt O15350). TP73 has no transmembrane segment (UniProt O15350 topology). The tissue profile is rated Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC).
| Distinct nuclear staining in bronchial or nasopharyngeal basal cells, or skin keratinocytes. | This matches the reported high staining in those cell populations (HPA tissue IHC). Assess the relevant cells individually: a convincing result need not color every cell in a section, because the reported tissue profile describes nuclear expression in subsets (HPA tissue IHC). |
| Predominantly diffuse cytoplasmic, membranous, or extracellular color, with little nuclear signal. | That is a poor match for the chiefly nuclear tissue pattern (HPA tissue IHC). UniProt also lists cytoplasm, so cytoplasmic signal alone cannot prove an artefact (UniProt O15350). Check whether the nuclear pattern appears in a known-positive control before interpreting the slide; if it does not, review detection and background using general IHC practice. |
| Strong staining in adipocytes or another cell population reported as undetected. | Adipocytes in adipose tissue are reported as not detected (HPA tissue IHC). Staining there warrants investigation of antibody cross-reactivity or endogenous detection activity using general IHC controls. A negative reference population is useful for assessing specificity, but one discrepant cell does not by itself identify the cause. |
| Broad color across epithelium, stroma, and empty spaces, without clear nuclear boundaries. | This cannot be scored confidently against the reported subset-specific nuclear pattern (HPA tissue IHC). In chromogenic IHC, inspect the no-primary control and review blocking, washes, and chromogen development as general workflow checks. Record the background separately from any discrete nuclear signal. |
| No nuclear staining in bronchial or nasopharyngeal basal cells, or in skin keratinocytes. | These are reported high-staining populations (HPA tissue IHC), so an entirely blank result needs an assay check before being called biological absence. Confirm that the expected cell population is present, then review the IHC-validated antibody, detection controls, and the assay's antigen-retrieval conditions as general IHC practice. |
| Cell population and tissue | High staining is reported in bronchial and nasopharyngeal basal cells and skin keratinocytes; medium staining in cervical, esophageal, and oral squamous epithelial cells (HPA tissue IHC). Compare like cell populations when judging intensity, since the supplied observations differ by tissue. |
| Predominant compartment | Nuclear expression in tissue is the principal IHC expectation (HPA tissue IHC). UniProt also records cytoplasm and nuclear accumulation after DNA damage (UniProt O15350); those annotations allow context-dependent interpretation, but do not validate a diffuse cytoplasmic tissue-IHC pattern. |
| Isoforms and antibody coverage | UniProt lists 11 TP73 isoforms, including transactivation-domain-containing and domain-lacking forms with different functions (UniProt O15350). Interpret a catalog antibody's result according to its documented epitope and isoform coverage; the supplied record gives no epitope or isoform coverage for that antibody. |
| Strength of IHC evidence | The tissue profile is Enhanced but has medium staining-to-RNA consistency (HPA tissue IHC). Two listed antibodies have Enhanced IHC status, CAB002514 and CAB003022 (HPA antibodies). That supports the reported pattern while leaving a new specimen's staining to be assessed with appropriate controls. |
| IF/ICC Q&A: should every labeled structure count? | No. Nucleoplasm is the supported main ICC-IF location; Golgi, vesicles, centrosome, and basal body are approved additional locations, while plasma membrane and cell junctions are uncertain (HPA subcellular). These cell-image observations do not establish the expected chromogenic tissue-IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive tissue is blank. | The expected basal-cell or keratinocyte population may be absent from the section, or the IHC assay may have failed; those populations are reported high (HPA tissue IHC). | Locate the relevant cells first. Then check the positive control, IHC-validated antibody instructions, retrieval conditions, and detection reagents as general IHC practice; do not infer TP73-specific fixation sensitivity from this record. |
| Signal appears only in cytoplasm. | UniProt lists cytoplasm, but the reported tissue-IHC profile is nuclear (UniProt O15350; HPA tissue IHC). Cytoplasm-only color therefore needs confirmation before scoring. | Compare a known-positive epithelial control for discrete nuclei and inspect a no-primary control. Review antibody and detection conditions if nuclear staining is also missing; these are general IHC checks. |
| Adipocytes stain strongly. | Adipocytes in adipose tissue are reported as not detected (HPA tissue IHC). Antibody cross-reactivity or endogenous detection activity are possible general IHC explanations. | Compare no-primary and appropriate detection controls, then reassess whether the color is cellular and reproducible. Treat the signal as unexplained until those checks support a specific interpretation. |
| The whole section has diffuse color. | Diffuse, poorly bounded color cannot be matched reliably to the subset-specific nuclear profile (HPA tissue IHC); excess background is a general IHC possibility. | Inspect a no-primary control, then review blocking, washing, antibody concentration, and chromogen development as general IHC practice. Re-score only distinct cellular staining after background is controlled. |
| Only a few cells stain in a positive tissue. | Nuclear expression is reported in subsets, and staining level varies among listed epithelial populations (HPA tissue IHC). A sparse pattern may reflect cell distribution or an assay issue. | Identify the cell type and compare its staining with the matching HPA tissue observation. If expected cells remain unstained, check the positive control and assay conditions before calling the sample negative. |
| An IF/ICC image seems to conflict with the IHC slide. | Nucleoplasm is the supported principal ICC-IF location, with several additional locations of differing support (HPA subcellular); tissue IHC reports a chiefly nuclear epithelial pattern (HPA tissue IHC). | Judge the paraffin-section result against tissue-IHC cell and nuclear patterns. Use the separate IF/ICC guide for that application; do not treat an additional ICC-IF location as proof of tissue-IHC positivity. |
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 |
|---|---|---|---|---|
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Skin | Keratinocytes | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | 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 TP73 staining in paraffin sections by checking retrieval, compartment, cell type and antibody epitope before interpreting chromogenic signal.
Catalog images show TP73 IHC in human skin cancer, human intestinal cancer and mouse brain, plus IF in 293 cells (catalog image captions). Listed reactivity spans human, mouse and rat (catalog reactivity).
M00688 has an IHC image from a human skin cancer paraffin section (M00688 IHC caption); PB9235 has IHC images from human intestinal cancer, rat brain and mouse intestine (PB9235 IHC captions). A00688-1 has an IHC image from formalin-fixed, paraffin-embedded mouse brain and an IF image from 293 cells (A00688-1 image captions).
Which to pick: For human paraffin-section IHC, choose M00688: its image caption specifies EDTA retrieval at pH 8.0 and a 1:50 primary dilution; the fixative is unreported (M00688 IHC caption). For IF/ICC, A00688-1 has an IF image in 293 cells (A00688-1 IF caption); M00688 also lists IF/ICC, but has no IF image in this payload (M00688 applications and image captions). For IHC across the listed human, mouse and rat species, choose PB9235, which has an IHC image for each; its captions do not report the fixative (PB9235 reactivity and IHC captions).