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ITPA / Inosine triphosphate pyrophosphatase · IHC design guide
Design Immunohistochemistry for ITPA
Plan ITPA staining in paraffin sections around the reported cytoplasmic tissue pattern (HPA tissue IHC). This guide pairs the catalog antibody’s 2–5 μg/ml IHC range (datasheet A02304) with positive and negative tissue controls drawn from the staining profile (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ITPA 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in multiple cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02304) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Esophagus+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (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 A02304) | |
| Caveat | Skin keratinocytes may be unstained despite broad expression (HPA tissue IHC) | |
| Regulation | No stimulus regulator specified (UniProt) | |
| Isoform / epitope | 3 isoforms; check whether the epitope is shared (UniProt) |
Section 1
Recommended ITPA IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A02304) is accompanied by published ITPA staining methods for mouse paraffin sections (PMC7710303) and renal carcinoma tissue microarrays (PMC5347726).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A02304) |
| Fixation | Image fixative and duration unreported (datasheet A02304); 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 A02304); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02304) |
| Primary antibody | Rabbit anti-ITPA, 2-5 μg/ml (datasheet A02304) |
| Primary incubation | Overnight at 4 °C (datasheet A02304) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02304) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ITPA-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet: A02304); citrate pH 6.0 pressure retrieval is a published alternative (PMC5347726).
Section 2
What Is the Expected ITPA Staining Pattern?
ITPA is cytoplasmic and has no transmembrane segment (UniProt Q9BY32). In paraffin sections, expect general cytoplasmic staining across cell types, with medium staining documented in selected adipocytes, glandular cells, respiratory epithelial cells and neuronal cells (HPA tissue IHC). HPA rates its tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression data (HPA tissue IHC).
| Cytoplasmic staining in adrenal glandular cells or bronchial respiratory epithelial cells. | This fits the reported general cytoplasmic pattern; both cell populations have medium staining (HPA tissue IHC). Score the identified cells and their cytoplasm, because nearby unstained structures do not change the recorded result for those cells (general IHC practice). |
| Predominantly membrane, extracellular or sharply nuclear staining in an IHC section. | Membrane or extracellular emphasis conflicts with cytoplasmic localisation and the absence of a transmembrane segment or signal peptide (UniProt Q9BY32; HPA tissue IHC). A nuclear IHC pattern also warrants review, but nuclear signal alone cannot be dismissed: HPA reports supported nucleoplasmic staining in ICC-IF (HPA subcellular). |
| Strong staining in skin keratinocytes or esophageal squamous epithelial cells. | Both populations are listed as not detected by tissue IHC (HPA tissue IHC). Review cell identification, the no-primary control and detection chemistry for cross-reactivity or endogenous activity (general IHC practice). A discordant result calls for investigation rather than an automatic biological conclusion. |
| Broad, even colour over cells and surrounding tissue, without clear cell boundaries. | This is difficult to score as cytoplasmic ITPA, whose tissue IHC profile is generally cytoplasmic (HPA tissue IHC). Compare a no-primary control and inspect washing, blocking and chromogen development; excess background can obscure a real cell-associated signal (general IHC practice). |
| No staining in a section expected to contain adrenal glandular cells. | Those cells have medium staining in the HPA tissue IHC record (HPA tissue IHC). Confirm their presence and assess section quality, retrieval, antibody dilution and detection controls before calling the sample negative (general IHC practice). HPA's Approved rating still carries only medium staining-to-RNA consistency (HPA tissue IHC). |
💡Expected ITPA appearanceA credible positive shows identifiable cytoplasmic colour, including medium staining in documented adrenal glandular or bronchial respiratory epithelial cells (HPA tissue IHC); dominant membrane or extracellular colour is discordant with ITPA localisation (UniProt Q9BY32; HPA tissue IHC).
| Tissue and cell selection | HPA reports medium staining in adipocytes, adrenal glandular cells, bronchial respiratory epithelial cells and selected neuronal cells; skin keratinocytes and esophageal squamous epithelial cells are not detected (HPA tissue IHC). Match the scored cell population to the record. |
| Compartment and topology | UniProt places ITPA in the cytoplasm and records no transmembrane segment or signal peptide (UniProt Q9BY32). These annotations support a cytoplasmic IHC expectation, without establishing how fixation or antigen retrieval affects staining. |
| Antibody evidence | HPA022824 is Approved for IHC; HPA073963 has no supplied IHC rating (HPA antibodies). HPA's tissue profile is Approved but has medium consistency with RNA expression data (HPA tissue IHC). Interpret unexpected patterns with that limit in view. |
| IF/ICC Q&A: Is nuclear signal expected? | Yes: HPA reports supported nucleoplasmic and cytosolic localisation in ICC-IF (HPA subcellular). That observation informs compartment interpretation; it does not establish an IHC staining intensity or an IF/ICC protocol. |
| Isoforms and epitope coverage | UniProt lists three ITPA isoforms (UniProt Q9BY32). The supplied records do not map the catalog antibody's epitope to those isoforms, so they cannot establish which variants its IHC staining detects. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue has no signal. | The expected adrenal glandular-cell staining is medium, not guaranteed in every section (HPA tissue IHC); a failed staining step is also possible (general IHC practice). | Verify the relevant cells are present, then check a documented positive section, retrieval, antibody dilution and detection reagents together (general IHC practice). |
| The whole section has diffuse brown colour. | Nonspecific background or endogenous detection activity can obscure cell boundaries (general IHC practice); this does not match HPA's general cytoplasmic profile (HPA tissue IHC). | Compare no-primary and detection controls, then review blocking, washing and chromogen development before scoring cells (general IHC practice). |
| Skin keratinocytes stain strongly. | That cell population is listed as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm the cells and compare control sections and a no-primary control. Record the discrepancy instead of treating the signal as established ITPA expression (general IHC practice; HPA tissue IHC). |
| Signal outlines membranes or lies outside cells. | This conflicts with cytoplasmic localisation and absent membrane-spanning or secretory features (UniProt Q9BY32; HPA tissue IHC). | Inspect section morphology and control staining, then reassess whether the apparent outline is background, an adjacent cell or a true intracellular signal (general IHC practice). |
| IHC shows prominent nuclear staining. | The tissue IHC profile is generally cytoplasmic, while ICC-IF supports nucleoplasmic as well as cytosolic localisation (HPA tissue IHC; HPA subcellular). | Document the compartment and compare controls and the cytoplasmic signal. Treat a nuclear-only IHC result as unresolved rather than automatically specific or artifactual (general IHC practice; HPA subcellular). |
| Staining differs across areas of one section. | Cell populations can have different recorded staining levels, including low or not-detected entries (HPA tissue IHC); uneven staining is also a technical possibility (general IHC practice). | Score each identified cell population separately and compare morphology and control staining across the section before assigning a tissue-wide result (general IHC practice). |
Experiment evidence
IHC & IF images showing ITPA staining
A02304 · IHCIHC analysis of ITPA using anti-ITPA antibody (A02304). ITPA was detected in a paraffin-embedded section of human colon adenocarcinoma tissu
A02304 · IHCIHC analysis of ITPA using anti-ITPA antibody (A02304). ITPA was detected in a paraffin-embedded section of human glioblastoma tissue. Heat
A02304 · IFIF analysis of ITPA using anti-ITPA antibody (A02304). ITPA was detected in a paraffin-embedded section of human intestinal cancer tissue. H
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000125877-ITPA/tissue/cerebral+cortex#imid_6205320
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000125877-ITPA/tissue/colon#imid_6203446
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000125877-ITPA/tissue/liver#imid_6203447
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000125877-ITPA/tissue/kidney#imid_6203456
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA022824; green = ITPA, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA022824; green = ITPA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ITPA IHC & IF
🧪Run adrenal gland first: its glandular cells should stain (HPA: Medium in adrenal gland glandular cells). Use salivary gland glandular cells as the tissue negative (HPA: Not detected); on the adrenal slide, unstained stromal cells can provide a background reference, but their ITPA status is not established by the supplied HPA row.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ITPA in HEL, U-251MG, U2OS, A-549, with annotated localisation: Nucleoplasm (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; nonimmune rabbit IgG matched to the primary antibody’s clonality where known (selected-SKU caption: rabbit primary); and an ITPA knockout biological negative. For adrenal gland IHC, quench endogenous peroxidase and check pigment or blood-associated background before scoring DAB staining (standard IHC practice).
⚠️Feasibility: The selected-SKU tissue-IHC caption reports heat retrieval in EDTA at pH 8.0 for a paraffin section, but reports no fixative; a target-specific fixation window or fixation effect is unreported (selected-SKU caption). Retrieval conditions for adrenal gland remain to be optimized rather than assumed from that caption. The supplied evidence does not establish whether frozen sections or IF are easier; HPA supports nucleoplasm and cytosol localization in ICC-IF images (HPA subcellular), while adrenal pigment and endogenous peroxidase warrant background checks in chromogenic IHC (standard IHC practice).
HPA tissue IHC evidence for ITPA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skin | Keratinocytes | Not detected | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ITPA IHC Tips
Troubleshoot ITPA staining in paraffin sections using the catalog antibody’s tissue image and independent localisation evidence (A02304 caption; HPA tissue IHC; HPA subcellular).
What retrieval should I try when ITPA staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02304). The catalog image used this retrieval before staining a human colon adenocarcinoma section with 2 μg/ml primary antibody overnight at 4°C (A02304 caption). If staining remains weak, compare a modest range of heating times on adjacent sections while keeping the buffer, antibody concentration and detection constant (standard IHC practice). Include a section processed without primary antibody to distinguish increased ITPA signal from retrieval related background (standard IHC practice). Record heating and cooling conditions so the comparison can be repeated (standard IHC practice).
Could fixation explain variable ITPA staining between paraffin blocks?
The catalog tissue caption identifies a paraffin embedded section but does not state its fixative or fixation time (A02304 caption). Target specific ITPA sensitivity to fixation is therefore unknown; do not infer it from the tissue staining profile or protein annotations (A02304 caption; HPA tissue IHC; UniProt Q9BY32). Compare sections with documented fixation histories using the same EDTA pH 8.0 retrieval and 2 μg/ml primary antibody conditions (datasheet A02304; standard IHC practice). Examine tissue preservation and staining together, since poorly preserved morphology makes a weak signal difficult to interpret (standard IHC practice). Keep section thickness and detection conditions consistent across the comparison (standard IHC practice).
How should I assess nuclear staining when ITPA is expected in cytoplasm?
Assess cytoplasmic staining first, because tissue IHC shows a general cytoplasmic pattern and UniProt places ITPA in the cytoplasm (HPA tissue IHC; UniProt Q9BY32). Nucleoplasmic staining can also be plausible: cell imaging supports both nucleoplasm and cytosol (HPA subcellular). Score nuclear and cytoplasmic signal separately in intact cells, using the counterstain to define nuclear boundaries (standard IHC practice). Compare each compartment with a no primary control and with nearby cells on the same section (standard IHC practice). A nuclear only DAB pattern should prompt a review of detection background and morphology before assignment to ITPA (HPA tissue IHC; HPA subcellular; standard IHC practice).
Can this stain distinguish the three ITPA isoforms?
ITPA has 3 annotated isoforms, but the supplied tissue caption does not identify the antibody epitope or establish isoform selectivity (UniProt Q9BY32; A02304 caption). Treat chromogenic staining as total detectable ITPA unless an epitope map and isoform specific validation become available (standard IHC interpretation). Check whether any proposed epitope is shared across isoforms before assigning a pattern to one transcript product (UniProt Q9BY32; standard antibody validation practice). The protein has no annotated transmembrane segment or glycosylation sites, and acetylation is reported near its N terminus; none of these annotations establishes this antibody’s epitope (UniProt Q9BY32). Keep retrieval and detection identical when comparing patterns across specimens (standard IHC practice).
How can IF help check which cells carry the ITPA signal?
Use an epithelial marker such as pan cytokeratin when examining the colon adenocarcinoma context of the catalog IHC image, and compare its cell boundaries with ITPA signal (A02304 caption; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence; a far red channel may improve contrast if shorter wavelengths are bright (standard IF practice). ITPA has no transmembrane segment and has supported cytosolic and nucleoplasmic locations, so permeabilise cells when the antibody must reach intracellular epitopes (UniProt Q9BY32; HPA subcellular; standard IF practice). Optimise IF fixation, permeabilisation and antibody concentration independently; the 2 μg/ml value comes from paraffin section IHC (A02304 caption; standard IF practice).
What should I check when DAB staining appears widespread?
Run a section without primary antibody through the same secondary and DAB steps to identify detection background (standard IHC practice). The catalog image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase linked secondary (A02304 caption). Block endogenous peroxidase before DAB development and compare staining in intact tissue with tissue edges or damaged regions (standard chromogenic IHC practice). If background persists, adjust blocking, washing and primary concentration one variable at a time while retaining the documented EDTA pH 8.0 retrieval as the reference condition (datasheet A02304; standard IHC practice).
How should I quantify ITPA staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and an intensity weighted H-score for cytoplasmic staining (HPA tissue IHC; standard IHC scoring practice). Score nucleoplasmic signal separately if present, because cell imaging supports that location too (HPA subcellular; standard IHC scoring practice). Normalise positive cell counts to the number of evaluable cells, or report positive cell density per mm² of viable annotated tissue (standard image analysis practice). Use the same DAB development, counterstain, imaging settings and positivity threshold across sections, and exclude folds and necrotic areas from the denominator (standard IHC quantification practice).
How do I separate credible ITPA staining from artefact?
A credible pattern should occupy intact cells and be compatible with the general cytoplasmic tissue profile; supported nucleoplasmic and cytosolic staining in cell imaging adds context for a nuclear component (HPA tissue IHC; HPA subcellular). Compare the cells of interest with expected variation: HPA reports medium staining in adipocytes and several glandular cell populations, but no detection in the listed skin keratinocytes (HPA tissue IHC). Treat signal concentrated at section edges, folds or necrotic areas as suspect, and check a no primary control for endogenous peroxidase or detection signal (standard IHC practice). Do not infer ITPA enzyme activity from DAB intensity alone (UniProt Q9BY32 function; standard IHC interpretation).
Boster reagents
Best ITPA / Inosine triphosphate pyrophosphatase IHC Antibodies
A02304 has documented paraffin-section IHC in human cancer tissues and IF in a human intestinal cancer section (catalog image captions); its listed reactivity includes human, mouse and rat (catalog applications/reactivity).
Real IHC data
Anti-ITPA Antibody ®
Cat # A02304
A02304 is the SKU with a rendered IHC figure, showing a human colon adenocarcinoma paraffin section; its other captions document human glioblastoma, liver cancer and lung cancer IHC, plus intestinal cancer IF (catalog image captions). M02304 lists human IHC, but has no IHC or IF image caption in the payload (catalog applications/reactivity; image captions).
Which to pick: Choose A02304 for paraffin-section IHC because its own caption documents that preparation and 2 μg/ml primary antibody (A02304 IHC caption); the fixative is unreported (A02304 IHC caption). For IF, choose A02304 based on its human intestinal cancer section image at 5 μg/ml; ICC evidence is unreported (A02304 IF caption). A02304 lists human, mouse and rat reactivity, while rabbit monoclonal M02304 lists human reactivity and IHC without a figure caption (catalog applications/reactivity; M02304 clone listing and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BY32 (ITPA_HUMAN, Inosine triphosphate pyrophosphatase).
- Human Protein Atlas. ITPA tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ITPA subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol..
- Human Protein Atlas. ITPA antibody validation summary (2 antibodies).
- Neural stem cell-specific ITPA deficiency causes neural depolarization and epilepsy. JCI insight 2020 — PMC7710303.
- Anti-diuretic hormone ITP signals via a guanylate cyclase receptor to modulate systemic homeostasis in Drosophila. eLife 2025 — PMC12611267.
- A four-gene signature predicts survival in clear-cell renal-cell carcinoma. Oncotarget 2016 — PMC5347726.
- First ever case report of co-occurrence of hobnail variant of papillary thyroid carcinoma and intrathyroid parathyroid adenoma in the same thyroid lobe. International journal of surgery case reports 2020 — PMC7218145.
- PubMed PMID:11278832 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ITPA (Inosine triphosphate pyrophosphatase). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02304). Validated IHC antibody: A02304 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Esophagus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A02304); 2. Fixation: Image fixative and duration unreported (datasheet A02304); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A02304); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02304); 8. Primary antibody: Rabbit anti-ITPA, 2-5 μg/ml (datasheet A02304); 9. Primary incubation: Overnight at 4 °C (datasheet A02304). Watch for: Fixation: Keep paraffin-section fixation consistent (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 A02304); Caveat: Skin keratinocytes may be unstained despite broad expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; check whether the epitope is shared (UniProt); Regulation: No stimulus regulator specified (UniProt). Evidence: UniProt Q9BY32, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol., Datasheet A02304, PMC7710303, PMC5347726. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ITPA/immunohistochemistry-ihc-guide.