TP73 / Tumor protein p73 · IHC design guide

Design Immunohistochemistry for TP73

Plan chromogenic TP73 IHC in paraffin sections using the nuclear staining reported in epithelial subsets as a reference (HPA tissue IHC). Check antibody epitope coverage when interpreting results across TP73’s 11 isoforms (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for TP73 (IHC for TP73): expected localisation Nuclear in stained epithelial subsets (HPA tissue IHC), antibody M00688, validated IHC image, and IHC protocol steps
Printable TP73 IHC protocol sheet — expected localisation Nuclear in stained epithelial subsets (HPA tissue IHC), antibody M00688, controls and protocol steps. Open the full TP73 IHC guide →

TP73 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 Nuclear in stained epithelial subsets (HPA tissue IHC)
Staining pattern Nuclear staining in subsets of glandular, squamous and respiratory epithelial 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
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific)
Caveat Isoform differences may affect antibody epitope coverage (UniProt)
Regulation Accumulates in nuclei after DNA damage (UniProt)
Isoform / epitope 11 isoforms; check which forms the antibody epitope covers (UniProt)
Section 1

Recommended TP73 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: M00688) is accompanied by published TP73 protocols for cervical cancer tissue (PMC6682548) and human liver tissue (PMC6920427).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human skin cancer tissue; fixative not specified (datasheet M00688)
FixationImage fixative and duration unreported (datasheet M00688); 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 M00688); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet M00688)
Primary antibodyRabbit monoclonal (clone EHI-20) anti-TP73, 1:50 (datasheet M00688)
Primary incubationOvernight at 4 °C (datasheet M00688)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet M00688)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultTP73-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.
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: M00688); use citrate pH 6.0 when reproducing the liver protocol (PMC6920427).
Section 2

What Is the Expected TP73 Staining Pattern?

TP73 is primarily nuclear in tissue IHC, with staining reported in subsets of respiratory and squamous epithelial cells (HPA: tissue IHC profile). Nucleoplasmic localization is supported by ICC-IF; UniProt also lists cytoplasm and nuclear accumulation after DNA damage (HPA: subcellular; UniProt O15350: subcellular location). TP73 has no transmembrane segment, so a membrane-only tissue pattern needs scrutiny (UniProt O15350: topology). HPA rates the tissue IHC pattern Enhanced, with medium consistency against RNA data (HPA: tissue IHC reliability).

What am I looking at on my slide?
Distinct nuclear staining in bronchial or nasopharyngeal basal cells, or skin keratinocytes, with limited surrounding signal (HPA: High in these cells).This fits the reported positive tissue pattern. Score the named cell population and nuclear compartment separately; a convincing section preserves recognizable morphology and a readable counterstain (HPA: tissue IHC; standard IHC practice).
Predominantly membrane, junctional, or diffuse cytoplasmic chromogen, with little nuclear signal.Do not score this alone as the expected IHC result: the tissue profile is nuclear (HPA: tissue IHC). UniProt permits cytoplasmic localization, and HPA reports additional ICC-IF locations, some uncertain; those observations do not validate a membrane-only tissue IHC pattern (UniProt O15350: subcellular location; HPA: subcellular).
Strong staining in adipocytes or in another cell population recorded as undetected in the selected HPA tissue.Consider cross-reactivity or endogenous detection activity, then inspect the exact tissue and cell annotation before calling it TP73. HPA lists adipocytes in adipose tissue as Not detected; its negatives are cell-specific, not proof that every cell in that tissue lacks TP73 (HPA: tissue IHC; standard IHC practice).
Haze across tissue and empty spaces, with nuclei and tissue boundaries hard to distinguish.Treat widespread diffuse color as background until controls support a cell-specific signal. Review blocking, antibody concentration, washing, and detection chemistry; a clean, localized nuclear pattern is the relevant comparator (HPA: tissue IHC profile; standard IHC practice).
No nuclear signal in an otherwise interpretable bronchus, nasopharynx, or skin section.A technical false negative is possible because HPA reports High staining in the specified basal cells or keratinocytes. Confirm that those cells are present, then check retrieval, antibody concentration, and detection controls before interpreting a negative result (HPA: tissue IHC; standard IHC practice).
💡Expected TP73 appearanceCall a positive result when discrete nuclear staining marks the appropriate epithelial cells—typically High in bronchial or nasopharyngeal basal cells or skin keratinocytes—while diffuse field-wide color or membrane-only staining remains suspect (HPA: tissue IHC; UniProt O15350: topology).
How each factor affects the staining
Tissue and cell selectionHPA reports High staining in bronchial and nasopharyngeal basal cells and skin keratinocytes; cervix, esophagus, and oral mucosa squamous cells are Medium. Choose and score the documented cell type, not the whole section (HPA: tissue IHC).
Compartment and assayNucleoplasm is the supported main ICC-IF location, whereas tissue IHC describes nuclear expression in selected epithelial cells. Additional ICC-IF locations do not establish a tissue IHC scoring pattern (HPA: subcellular; HPA: tissue IHC).
Isoforms and response stateUniProt lists 11 isoforms and nuclear accumulation after DNA damage. An isoform-specific or damage-response claim needs epitope and sample evidence beyond this tissue pattern; avoid inferring either from staining intensity alone (UniProt O15350: isoforms and subcellular location).
Antibody validationThe HPA tissue pattern is Enhanced, and two listed antibodies have Enhanced IHC status; HPA also reports medium consistency with RNA. This supports the reported pattern without proving that every reagent or specimen will reproduce it (HPA: tissue IHC reliability; HPA: antibody validation).
Antigen retrieval and detectionUse the catalog antibody's IHC-P instructions to set retrieval and dilution, then assess nuclear signal against background (standard IHC practice). No TP73-specific fixation sensitivity or retrieval effect is established by the supplied UniProt and HPA records.
IF/ICC Q&AQ: Should IF puncta or membrane signal be scored as the IHC result? A: No; HPA supports nucleoplasm as the main ICC-IF location and marks plasma membrane and junctions uncertain. Evaluate IF/ICC on its own guide page (HPA: subcellular).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Positive-control section has no nuclear staining.The expected basal cells or keratinocytes may be absent, or retrieval, antibody application, or detection may have failed (HPA: High in named cells; standard IHC practice).Verify the annotated cell population on the slide; check the IHC-P instructions and detection controls, then repeat with a documented positive section (HPA: tissue IHC; standard IHC practice).
Color covers most of the section.Nonspecific binding, excess antibody, inadequate washing, or detection background can obscure the expected nuclear distribution (HPA: tissue IHC profile; standard IHC practice).Review blocking and washes, titrate within the catalog IHC-P guidance, and compare a no-primary control with the stained section (standard IHC practice).
Color appears mainly at membranes or junctions.This differs from HPA's nuclear tissue IHC profile; HPA labels membrane and junctional ICC-IF locations uncertain, so staining identity is unresolved (HPA: tissue IHC; HPA: subcellular).Check cellular morphology and controls, then seek reproducible nuclear staining in a documented positive cell type before scoring the IHC result (HPA: tissue IHC; standard IHC practice).
An HPA-listed negative cell type stains strongly.Cross-reactivity or endogenous detection activity is possible; the HPA negative entry applies to the specified cell type, such as adipocytes in adipose tissue (HPA: tissue IHC; standard IHC practice).Confirm cell identity and review no-primary and detection controls; avoid extending the HPA cell-specific negative call to the entire tissue (HPA: tissue IHC; standard IHC practice).
Fallopian tube staining seems mixed or hard to score.HPA's profile mentions nuclear staining in a subset of glandular cells, while its cell table records Low staining in ciliated cell bodies; these descriptions warrant cell-level review (HPA: tissue IHC).Record the cell type and compartment seen, and use a High, clearly annotated tissue as the positive comparator rather than forcing one fallopian tube-wide score (HPA: tissue IHC; standard IHC practice).
Nuclear staining varies between specimens.TP73 can accumulate in nuclei after DNA damage, while HPA reports only medium consistency between antibody staining and RNA; these facts alone do not establish why a given specimen differs (UniProt O15350: subcellular location; HPA: tissue IHC reliability).Compare like cell populations, morphology, and run controls; report the observed fraction and intensity without assigning a damage state or isoform from IHC alone (HPA: tissue IHC; UniProt O15350: isoforms; standard IHC practice).

Sample controls for TP73 IHC & IF

🧪Run bronchus first and look for staining in basal cells (HPA: High in bronchus basal cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the bronchus slide, adjacent cells without specific signal should lack nuclear chromogen, with diffuse color treated as background (standard IHC practice).
Positive control tissue: Bronchus (Basal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TP73 in HEK293, Rh30, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the rabbit primary antibody and its clonality, and a TP73-knockout biological control (caption: rabbit anti-TP73; standard IHC practice). Block endogenous peroxidase and check inflammatory cells for residual chromogen in bronchus sections (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M00688 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0 and a 1:50 primary dilution overnight at 4°C, but does not establish that retrieval is required (caption: M00688 tissue IHC). Frozen sections are not established as easier; ICC-IF images exist, but their availability does not establish an easier tissue workflow (HPA: ICC-IF cell-line images).

HPA tissue IHC evidence for TP73

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced TP73 IHC Tips

Troubleshoot TP73 staining in paraffin sections by checking retrieval, cell type, nuclear localisation and antibody epitope before interpreting chromogenic signal.

Which retrieval condition should I try first for TP73 in paraffin sections?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M00688). The selected tissue image used this condition before overnight incubation with the catalog antibody at 1:50 and 4°C (caption M00688). If nuclear staining is weak, check that heating, cooling and buffer preparation are consistent across the run before changing retrieval conditions (standard IHC practice). Compare any fallback buffer or pH on adjacent sections, keeping antibody dilution and DAB development constant (standard IHC practice). Judge recovery in epithelial nuclei, since nuclear staining is the principal tissue pattern reported for TP73 (HPA tissue IHC).
Could fixation explain weak or uneven TP73 staining?
The selected image shows a paraffin section, but its fixative is unreported; TP73-specific sensitivity to fixation is therefore unknown (caption M00688). Record the fixative and fixation duration for each specimen, and compare sections with matched processing when investigating uneven signal (standard IHC practice). Check section adhesion and morphology after heat retrieval, because tissue loss or damage can resemble patchy antigen loss (standard IHC practice). Keep retrieval at EDTA pH 8.0 while assessing processing differences (datasheet M00688). Use a concurrently stained section containing expected positive epithelial cells to distinguish a processing problem from genuinely sparse expression (HPA tissue IHC; standard IHC practice).
Should TP73 stain nuclei, cytoplasm or both?
Prioritise nuclear staining when reading chromogenic TP73 sections: the tissue profile reports nuclear expression in selected epithelial cells (HPA tissue IHC). TP73 is also annotated in the cytoplasm, and can accumulate in the nucleus after DNA damage (UniProt O15350 localisation). If cytoplasmic DAB dominates, inspect nuclear counterstaining, cell boundaries and a no-primary control before assigning that pattern to TP73 (standard IHC practice). Compare staining within the expected cell population rather than treating every stained compartment as equivalent (HPA tissue IHC; standard IHC practice). Document nuclear and cytoplasmic results separately so a shift in localisation remains visible during review (UniProt O15350 localisation; standard IHC practice).
Can this antibody distinguish TP73 isoforms or modified epitopes?
Do not assign an isoform from a positive DAB signal unless the antibody epitope and isoform coverage are established (standard IHC practice). TP73 has 11 listed isoforms, including forms with different transactivation domains, so shared epitopes can obscure biologically distinct products (UniProt O15350 isoforms and function). Its SAM domain spans residues 485–551, while reported modified residues include 27, 28 and 99 (UniProt O15350 domains and modified residues). Check the catalog antibody's immunogen information before claiming recognition of a particular region or modification (standard IHC practice). Record exactly which isoform or phospho-specific inference the available epitope information supports; otherwise report TP73 immunoreactivity (standard IHC practice).
How should I plan an IF follow-up to a TP73 IHC result?
Treat IF/ICC as a separate assay when following up a chromogenic TP73 result; the selected product evidence describes paraffin-section IHC (caption M00688). Multiplex TP73 with a marker for the expected epithelial cell population, choosing the marker from the sampled tissue and validating channel separation (HPA tissue IHC; standard IF practice). Select fluorophores after checking tissue autofluorescence and include single-label and no-primary controls for spectral bleed-through and background (standard IF practice). Because TP73 has no transmembrane segment and its main supported location is nucleoplasmic, validate permeabilisation that gives antibody access to nuclear epitopes (UniProt O15350 topology; HPA subcellular). Compare nuclear signals at matched acquisition settings (standard IF practice).
How can I reduce diffuse or misleading DAB background?
Start with a no-primary control and inspect whether colour follows cells, tissue edges or damaged areas (standard IHC practice). The selected image used 10% goat serum blocking, a peroxidase-conjugated secondary and DAB detection; its primary incubation was 1:50 overnight at 4°C (caption M00688). Include an endogenous peroxidase block and control DAB development as general chromogenic IHC steps (standard IHC practice). If background persists, verify secondary-antibody specificity and compare a lower primary concentration on adjacent sections while keeping retrieval fixed (standard IHC practice). Evaluate improvement in expected epithelial nuclei, where TP73 tissue staining is reported (HPA tissue IHC).
How should I score heterogeneous TP73 staining in tissue? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population before measuring TP73, since the reported tissue pattern is restricted to subsets of epithelial cells (HPA tissue IHC). For nuclear DAB, record the percentage of positive cells and intensity, or calculate an H-score using consistent intensity bins (standard IHC practice). Normalise counts to the number of eligible cells; if reporting positive-cell density, state the sampled area in mm² (standard IHC practice). Score nuclear and cytoplasmic staining separately, because TP73 has both annotated locations but the reported tissue pattern is nuclear (UniProt O15350 localisation; HPA tissue IHC). Keep exposure, counterstain assessment and scoring thresholds consistent across compared sections (standard IHC practice).
What makes a TP73-positive IHC result convincing?
Look for reproducible nuclear staining in the appropriate cells: TP73 is reported in epithelial nuclei, with high staining in skin keratinocytes and bronchial basal cells (HPA tissue IHC). Compare that pattern with a no-primary control and with morphology on the counterstained section (standard IHC practice). Signal confined to tissue edges, necrotic areas or regions showing endogenous peroxidase activity warrants review before scoring (standard IHC practice). Dominant staining in an unexpected compartment or cell population needs independent validation, since TP73 also has an annotated cytoplasmic location (UniProt O15350 localisation; standard IHC practice). Interpret a negative result cautiously because the HPA tissue profile reports medium agreement between staining and RNA expression (HPA tissue IHC).
Boster reagents

Best TP73 / Tumor protein p73 IHC Antibodies

Explore anti-TP73 antibodies with IHC images from human skin and intestinal cancers, mouse brain and intestine, and rat brain, plus a 293-cell IF image (catalog image captions).

Real IHC data IHC analysis of TP73 using anti-TP73 antibody (M00688). TP73 was detected in a paraffin-embedded section of human skin 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 1:50 rabbit anti-TP73 Antibody (M00688) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-p73 Rabbit Monoclonal Antibody
Cat # M00688
Real IHC data Anti-p73 antibody, PB9235, IHC(P) IHC(P): Human Intestinal Cancer Tissue
Anti-p73/TP73 Antibody ®
Cat # PB9235
Real IHC data TP73 Antibody (Center) IHC analysis in formalin fixed and paraffin embedded mouse brain followed by peroxidase conjugation of the secondary antibody and DAB staining. This data demonstrates the use of the TP73 Antibody (Center) for immunohistochemistry. Clinical relevance has not been evaluated.
Anti-TP73 Antibody (Center)
Cat # A00688-1

M00688 has a paraffin-section IHC image of human skin cancer, while PB9235 has IHC(P) images of human intestinal cancer, mouse intestine, and rat brain (catalog: M00688 and PB9235 IHC captions). A00688-1 has an IHC image of formalin-fixed, paraffin-embedded mouse brain and an IF image of 293 cells (catalog: A00688-1 IHC and IF captions).

Which to pick: For human tissue IHC, M00688 is a rabbit monoclonal shown on a paraffin section of human skin cancer at 1:50; its IHC caption does not report a fixative (catalog: M00688 host, clone, reactivity, and IHC caption). For IF/ICC, M00688 lists both applications for human samples at 1:50; A00688-1 lists IF for human and mouse at 1:10–1:50 and has a 293-cell IF image (catalog: M00688 and A00688-1 applications, reactivity, and IF dilutions; A00688-1 IF caption). For tissue IHC across human, mouse, and rat, PB9235 has IHC(P) images from all three species; its captions do not report a fixative (catalog: PB9235 reactivity and IHC captions).

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

References

  1. UniProt Consortium. UniProt entry O15350 (P73_HUMAN, Tumor protein p73).
  2. Human Protein Atlas. TP73 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. TP73 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane, cell junctions, the Golgi apparatus, vesicles, centrosome and basal body..
  4. Human Protein Atlas. TP73 antibody validation summary (4 antibodies).
  5. TP73 is a credible biomarker for predicting clinical progression and prognosis in cervical cancer patients. Bioscience reports 2019 — PMC6682548.
  6. DNA Methylation Activates TP73 Expression in Hepatocellular Carcinoma and Gastrointestinal Cancer. Scientific reports 2019 — PMC6920427.
  7. Profiling hypoxia signaling reveals a lncRNA signature contributing to immunosuppression in high-grade glioma. Frontiers in immunology 2024 — PMC11479907.
  8. Distinct p63 and p73 Protein Interactions Predict Specific Functions in mRNA Splicing and Polyploidy Control in Epithelia. Cells 2020 — PMC7824480.
  9. PubMed PMID:9288759 — UniProt-cited evidence.
  10. PubMed PMID:9721206 — UniProt-cited evidence.
  11. PubMed PMID:9802988 — UniProt-cited evidence.