TP73 / Tumor protein p73 · IHC design guide

Design Immunohistochemistry for TP73

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).

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 tissue; cytoplasmic location also annotated (HPA tissue IHC: nuclear; UniProt: cytoplasm), antibody M00688, validated IHC image, and IHC protocol steps
Printable TP73 IHC protocol sheet — expected localisation Nuclear in tissue; cytoplasmic location also annotated (HPA tissue IHC: nuclear; UniProt: cytoplasm), 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended TP73 IHC & IF Protocols

The catalog antibody uses EDTA pH 8.0 retrieval (datasheet M00688). Published IHC protocols cover cervical cancer and liver sections (PMC6682548; 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 heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet M00688); compare the published retrieval conditions during optimization (PMC6682548; PMC6920427).
Section 2

What Is the Expected TP73 Staining Pattern?

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).

What am I looking at on my slide?
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.
💡Expected TP73 appearanceCall a result positive when distinct nuclear color marks the expected epithelial cells—often high in bronchial or nasopharyngeal basal cells or skin keratinocytes (HPA tissue IHC); widespread, compartment-free color is suspect by general IHC practice.
How each factor affects the staining
Cell population and tissueHigh 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 compartmentNuclear 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 coverageUniProt 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 evidenceThe 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.
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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.

Sample controls for TP73 IHC & IF

🧪Run skin first and expect staining in keratinocytes (HPA: High in skin keratinocytes). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the skin slide, assess non-keratinocyte cells for background staining without assuming they are TP73-negative (HPA: skin result specifies keratinocytes).
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 no-primary (secondary-only) and rabbit IgG isotype controls matched to the primary antibody’s class and clonality, plus a TP73-knockout biological control (selected IHC caption: rabbit anti-TP73 M00688). Quench endogenous peroxidase for the HRP/DAB assay, and distinguish skin pigment from chromogen (selected IHC caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: The selected skin-cancer paraffin-section caption reports heat-mediated retrieval in EDTA at pH 8.0, providing a starting condition for IHC-P (selected IHC caption: EDTA pH 8.0); its fixative is unreported, and no target-specific fixation window or fixation effect is reported in the supplied evidence. The supplied evidence does not establish that frozen sections or IF/ICC are easier for TP73, while skin pigment may complicate DAB interpretation (selected IHC caption: paraffin-section HRP/DAB; standard IHC practice).

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, compartment, cell type and antibody epitope before interpreting chromogenic signal.

How should I retrieve TP73 in paraffin sections when nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M00688). The selected paraffin-section image used that retrieval before overnight incubation with the catalog antibody at 1:50 and 4°C (datasheet M00688). If nuclear staining remains weak, compare retrieval duration on adjacent sections while holding antibody dilution and DAB development constant, and inspect tissue morphology for heat damage (standard IHC practice). Include a skin section with keratinocytes as a reference, since those cells show high TP73 staining, but treat that reference as a staining check rather than proof of specificity (HPA: high in skin keratinocytes; standard IHC practice).
Could fixation explain weak or uneven TP73 staining in my paraffin sections?
TP73-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (datasheet M00688). Record each specimen’s fixative, fixation interval and processing history before comparing staining across cases (standard IHC practice). Compare adjacent sections processed together, using the reported EDTA pH 8.0 retrieval and 1:50 primary incubation as a starting point (datasheet M00688; standard IHC practice). If a processing batch stains unevenly, assess morphology and staining in an on-slide reference tissue before attributing a negative nucleus to absent TP73 (standard IHC practice). Do not infer fixation tolerance from tissue expression or protein features (HPA: tissue IHC profile; UniProt O15350: TP73 record).
How should I assess cytoplasmic TP73 staining when nuclear staining is expected?
Score nuclear and cytoplasmic DAB separately: TP73 is reported in both compartments, and nuclear accumulation can follow DNA damage (UniProt O15350: subcellular location). Nuclear staining has an independent tissue reference in subsets of glandular cells and in squamous and respiratory epithelium (HPA: tissue IHC profile). If only diffuse cytoplasmic color appears, examine a section processed without primary antibody and compare cells within the same tissue for nonspecific deposition (standard IHC practice). TP73 has no transmembrane segment, so a membrane-only pattern warrants additional validation rather than an automatic positive call (UniProt O15350: topology; standard IHC practice). Report compartment and cell type alongside staining intensity (standard IHC practice).
Can a negative TP73 stain exclude all TP73 isoforms?
A negative section cannot exclude every TP73 product: the record lists 11 isoforms, including Alpha, Beta, Gamma and dN forms (UniProt O15350: isoforms). First verify which sequence the catalog antibody recognizes and whether that sequence is retained in the isoforms relevant to the study; the supplied caption gives no epitope map (datasheet M00688; standard IHC practice). Compare staining with an independently characterized antibody against a distinct epitope when isoform coverage matters (standard IHC practice). Keep nuclear and cytoplasmic results separate because TP73 can occupy both compartments (UniProt O15350: subcellular location). Avoid assigning a DAB signal to a transactivating or dN isoform without isoform-specific validation (UniProt O15350: function and isoforms; standard IHC practice).
How can IF help resolve ambiguous TP73 staining in paraffin sections?
For a separate IF assay, pair TP73 with a marker that identifies the cell population under study, such as an epithelial marker when examining the reported squamous or respiratory epithelial pattern (HPA: tissue IHC profile; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence in each channel, and include single-label controls when signals will be merged (standard IF practice). TP73 has no transmembrane segment and is reported in the nucleus and cytoplasm, so permeabilize cells when the antibody must reach an intracellular epitope (UniProt O15350: topology and subcellular location; standard IF practice). Verify the antibody’s IF suitability and epitope accessibility independently; the selected M00688 caption establishes a paraffin-section chromogenic workflow only (datasheet M00688).
What should I check when TP73 DAB staining is widespread or granular?
Check a section processed without primary antibody to distinguish antibody-dependent staining from chromogen or detection background (standard IHC practice). The selected paraffin-section workflow used 10% goat serum, a peroxidase-conjugated secondary antibody and DAB, so match those steps when troubleshooting the pictured result (datasheet M00688). Include a peroxidase-blocking step and inspect wash quality if DAB appears broadly deposited; these are general chromogenic IHC controls, not TP73-specific requirements (standard IHC practice). Compare the pattern with the reported nuclear expression in subsets of epithelial cells and with high staining in skin keratinocytes (HPA: tissue IHC profile). Recheck sections showing pigment, folds or damaged edges before scoring them (standard IHC practice).
How should I quantify TP73 IHC across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measurement, then report nuclear percent-positive cells and an intensity-weighted H-score if staining varies across cells (standard IHC practice). Count within comparable tissue regions and normalize positive-cell counts to all eligible cells of that population, rather than to the whole section (standard IHC practice). If spatial abundance is the endpoint, report positive-cell density per mm² of viable, annotated tissue and document excluded regions (standard IHC practice). Record cytoplasmic staining separately because TP73 is reported in both nucleus and cytoplasm (UniProt O15350: subcellular location). Use consistent thresholds and an on-slide reference when comparing batches (standard IHC practice).
How do I distinguish convincing TP73 positivity from tissue or detection artefacts?
A convincing call combines staining in the expected cell population with a plausible compartment: TP73 is reported in nucleus and cytoplasm, and HPA describes nuclear expression in selected epithelial cells (UniProt O15350: subcellular location; HPA: tissue IHC profile). Check skin keratinocytes as a reference population, while recognizing that their high reported staining does not establish specificity in another specimen (HPA: high in skin keratinocytes; standard IHC practice). Discount isolated staining at section edges, folds or necrotic regions until it is reproduced in intact tissue (standard IHC practice). Use a no-primary control and peroxidase blocking to evaluate endogenous enzyme and DAB-related color (standard IHC practice). Treat an unexpected cell type or membrane-only pattern as a reason for independent validation (UniProt O15350: topology; standard IHC practice).
Boster reagents

Best TP73 / Tumor protein p73 IHC Antibodies

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).

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 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).

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.