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ITPKC / Inositol-trisphosphate 3-kinase C · IHC design guide
Design Immunohistochemistry for ITPKC
Plan paraffin-section ITPKC staining around cytoplasmic and nuclear signal (HPA tissue IHC). Use medium-staining adrenal glandular cells as a reference (HPA tissue IHC), and start the catalog antibody at 1:100–1:300 (datasheet A30549).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ITPKC 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 | Cytoplasmic and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic and nuclear staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30549) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
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 | High pancreatic expression, but low exocrine-cell staining (UniProt; HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one chain with no transmembrane segment (UniProt) |
Section 1
Recommended ITPKC IHC & IF Protocols
The catalog antibody IHC-P protocol (datasheet: A30549) is paired with a published cerebral cortex IHC protocol (PMC13343476).
| Sample | Paraffin-embedded Human lung cancer tissue; fixative not specified (datasheet A30549) |
| Fixation | Image fixative and duration unreported (datasheet A30549); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0 (datasheet A30549); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-ITPKC, 1:100 - 1:300 (datasheet A30549) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ITPKC-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 8.0 (datasheet: A30549); the published protocol does not specify retrieval (PMC13343476).
Section 2
What Is the Expected ITPKC Staining Pattern?
ITPKC can localise to both nucleus and cytoplasm and actively shuttles between them; it has no transmembrane segment (UniProt Q96DU7). In paraffin-section IHC, expect cell-associated nuclear and cytoplasmic staining, including medium staining in appendix glandular cells, bone marrow hematopoietic cells, and cerebellar Purkinje cells (HPA tissue IHC). HPA calls the tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Distinct nuclear and cytoplasmic chromogen in appendix glandular cells, with medium intensity (HPA tissue IHC). | This fits the reported cell type and intensity. Both compartments are plausible because ITPKC shuttles between nucleus and cytoplasm (UniProt Q96DU7). Judge the pattern against neighboring tissue and the negative control; brown color alone does not identify its source (general IHC practice). |
| Staining is restricted to membranes or extracellular material, with no convincing cellular nuclear or cytoplasmic signal. | This does not fit the reported general nuclear and cytoplasmic profile (HPA tissue IHC). ITPKC has no transmembrane segment or annotated signal peptide (UniProt Q96DU7). Check morphology, the negative control, and whether chromogen has deposited outside cells before assigning a compartment (general IHC practice). |
| The strongest staining is in a different cell population from the reported positive cells in the same tissue. | For example, HPA reports medium staining in bronchial respiratory epithelial cells (HPA tissue IHC). An unexpected dominant pattern warrants checking cell identification, nonspecific antibody binding, and endogenous detection activity (general IHC practice). HPA's broad tissue profile does not establish that every other cell must be negative (HPA tissue IHC). |
| Brown haze covers stroma and cells without discernible cellular boundaries or nuclear detail. | Diffuse deposition cannot establish ITPKC localisation; the reference profile is cellular, with general nuclear and cytoplasmic expression (HPA tissue IHC). Review the negative control and detection background, then assess whether blocking, washing, or chromogen development needs adjustment (general IHC practice). |
| No signal appears in an appendix section expected to contain glandular cells with medium staining (HPA tissue IHC). | First confirm that the relevant cells are present and that the positive-control run developed (general IHC practice). A blank slide may reflect an assay failure or an unsuitable section; it does not, by itself, overturn the reported HPA observation (HPA tissue IHC). |
💡Expected ITPKC appearanceCall a positive result when identifiable cells show convincing nuclear and/or cytoplasmic staining at the expected medium level in an HPA-reported population; isolated extracellular color or diffuse haze is insufficient (HPA tissue IHC; general IHC practice).
| Compartment readout | HPA tissue IHC describes general cytoplasmic and nuclear expression, consistent with active shuttling in the UniProt record (HPA tissue IHC; UniProt Q96DU7). HPA's supported nuclear-speckle localisation comes from ICC-IF; routine chromogenic IHC need not resolve speckles (HPA subcellular ICC-IF; general IHC practice). |
| Choice of reference cells | HPA reports medium staining in appendix glandular cells, bone marrow hematopoietic cells, and cerebellar Purkinje cells, but low staining in pancreatic exocrine glandular cells and liver cholangiocytes (HPA tissue IHC). Select the reference by its annotated cell population, not solely by the organ name (HPA tissue IHC). |
| Tissue-level expression versus cellular staining | UniProt describes high expression in pancreas and liver, while HPA records low IHC staining in the specified pancreatic exocrine cells and liver cholangiocytes (UniProt Q96DU7; HPA tissue IHC). These statements address different measurements or cell populations; neither predicts strong staining in those named cells (UniProt Q96DU7; HPA tissue IHC). |
| Strength of the IHC evidence | The HPA tissue profile is Approved with medium staining–RNA consistency; both listed rabbit polyclonal antibodies have Approved IHC status (HPA tissue IHC; HPA antibodies). This supports a reference pattern but leaves an unexpected specimen-specific result to be evaluated with controls and morphology (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive appendix glandular cells remain blank (HPA tissue IHC). | The expected cells may be absent from the section, or the IHC run may have failed (general IHC practice). | Confirm cell identity and section quality; check the concurrent positive control, then review retrieval and detection steps used for the validated IHC assay (general IHC practice). No ITPKC-specific retrieval condition is supplied. |
| Signal appears only at membranes or outside cells. | That location conflicts with the reported nuclear and cytoplasmic pattern and the absence of a transmembrane segment (HPA tissue IHC; UniProt Q96DU7). | Inspect the negative control and tissue morphology, and repeat scoring using identifiable cells and compartments (general IHC practice). Do not count isolated extracellular chromogen as a positive cell. |
| A broad brown haze obscures the counterstain. | Nonspecific binding, endogenous detection activity, or excessive chromogen development can produce background in chromogenic IHC (general IHC practice). | Compare with the negative control; review blocking, washes, detection reagents, and development time under the laboratory's IHC workflow (general IHC practice). |
| Unexpected cells stain more strongly than the HPA-reported population. | Cell misidentification, cross-reactivity, or endogenous detection activity are possible explanations (general IHC practice). | Recheck morphology and controls, then compare the exact tissue and cell population with the HPA profile. Treat an unlisted cell as unresolved rather than automatically negative (HPA tissue IHC; general IHC practice). |
| Pancreatic exocrine cells or liver cholangiocytes give only weak signal. | HPA records low IHC staining in those cells, although UniProt describes high expression in pancreas and liver (HPA tissue IHC; UniProt Q96DU7). | Score the specified cells against their HPA IHC levels; use a reported medium-staining cell population when assessing assay performance (HPA tissue IHC; general IHC practice). |
| IF/ICC: should a positive result show nuclear speckles? | HPA supports nuclear-speckle localisation in ICC-IF images, whereas tissue IHC has a broader nuclear and cytoplasmic profile (HPA subcellular ICC-IF; HPA tissue IHC). | For IF/ICC interpretation, compare nuclear signal with the supported speckle pattern (HPA subcellular ICC-IF). Assess chromogenic tissue IHC by its reported cellular pattern without requiring visible speckles (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ITPKC staining
A30549 · IHCImmunohistochemical analysis of paraffin-embedded Human lung cancer. Antibody was diluted at 1:100(4° overnight). High-pressure and temperat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000086544-ITPKC/tissue/cerebral+cortex#imid_14907432
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000086544-ITPKC/tissue/colon#imid_14907358
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000086544-ITPKC/tissue/liver#imid_14907359
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000086544-ITPKC/tissue/kidney#imid_14907344
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA053003; green = ITPKC, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA053003; green = ITPKC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ITPKC IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain at the HPA Medium level (HPA: bone marrow, hematopoietic cells, Medium). HPA detects ITPKC in all 44 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect anucleate erythrocytes on the positive slide to show only background after appropriate blocking (HPA: no negative tissues; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: None in HPA: ITPKC is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ITPKC in A-431, RT-4, U2OS, with annotated localisation: Nuclear speckles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype control matched to the primary antibody’s host species and immunoglobulin class; add ITPKC-knockout tissue if available, or a peptide-block control as shown in the selected caption (caption: peptide-preabsorbed negative control). In bone marrow, block endogenous peroxidase and assess residual signal in hematopoietic cells before interpreting chromogenic staining (HPA: bone marrow, hematopoietic cells; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A30549 paraffin-section caption does not state a fixative (caption: fixative unreported). That caption uses high-pressure, high-temperature Tris-EDTA pH 8.0 retrieval, but does not establish that this retrieval is required for all specimens (caption: retrieval conditions). The supplied evidence does not establish whether frozen sections or IF are easier; for bone marrow IHC, endogenous peroxidase is a practical source of false-positive chromogen (HPA: bone marrow, hematopoietic cells; standard IHC practice).
HPA tissue IHC evidence for ITPKC
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic 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 |
|---|---|---|---|---|
| None in HPA: ITPKC is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ITPKC IHC Tips
Troubleshoot ITPKC staining in paraffin sections by checking retrieval, controls, cell identity and compartment before interpreting chromogenic signal.
How should I retrieve ITPKC antigen when staining paraffin sections?
Use heat-mediated Tris-EDTA at pH 8.0 for antigen retrieval (datasheet A30549). The paraffin-section image used high pressure and temperature retrieval, followed by antibody at 1:100 overnight at 4 °C (caption A30549). If signal is weak, adjust heating and cooling consistently across matched sections while checking that nuclei and tissue architecture remain intact (standard IHC practice). Compare each change with a no-primary control and a section stained using the documented method (standard IHC practice). The caption also reports loss of staining after immunogen-peptide preabsorption, which provides a useful comparison for questionable signal (caption A30549).
Could fixation explain variable ITPKC staining across paraffin blocks?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (caption A30549). Record each block’s fixation history and compare blocks processed alike before attributing staining differences to ITPKC abundance (standard IHC practice). Apply the documented Tris-EDTA retrieval at pH 8.0 and hold antibody dilution, incubation and detection constant across the comparison (datasheet A30549; standard IHC practice). If fixation histories differ, assess morphology and control staining alongside ITPKC, then report the comparison as potentially affected by processing (standard IHC practice). Nuclear and cytoplasmic expression does not establish how this antigen responds to fixation (UniProt Q96DU7).
Should I score nuclear and cytoplasmic ITPKC staining separately?
Yes: ITPKC shuttles between nucleus and cytoplasm, so record staining in each compartment separately (UniProt Q96DU7). Tissue IHC shows general cytoplasmic and nuclear expression, while cell-based IF supports a nuclear-speckle location (HPA tissue IHC; HPA subcellular). Assess compartments within identified cells instead of treating every dark nucleus or diffuse cytoplasmic deposit as equivalent signal (standard IHC practice). Use a nuclear counterstain and inspect serial sections with consistent exposure to chromogen before scoring compartment patterns (standard IHC practice). Because tissue IHC reliability is Approved with medium staining–RNA consistency, describe compartment shifts cautiously and retain the underlying images for review (HPA tissue IHC).
How can epitope uncertainty affect interpretation of ITPKC staining?
The record lists one chain spanning residues 1–683 and no annotated isoforms, but the supplied caption does not map this antibody’s epitope (UniProt Q96DU7; caption A30549). Do not assign a weakly stained compartment to an alternative isoform without independent evidence (UniProt Q96DU7; standard IHC practice). Annotated phosphorylation sites include serine 127, serine 162, threonine 336 and serine 404; their effect on this antibody’s staining is unreported (UniProt Q96DU7; caption A30549). If staining varies, compare the same antibody and retrieval conditions across matched sections, and use the reported peptide-preabsorption result as context for specificity (caption A30549; standard IHC practice).
How should I investigate ITPKC localisation with multiplex immunofluorescence?
For an IF follow-up, pair ITPKC with a cell-identity marker, such as a standard epithelial marker when examining bronchial respiratory epithelium (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel where tissue autofluorescence interferes with shorter-wavelength signal (standard IF practice). Permeabilise sufficiently for intracellular access: ITPKC is nuclear and cytoplasmic and has no annotated transmembrane segment, although this antibody’s epitope is unmapped (UniProt Q96DU7; caption A30549). Include single-stain and no-primary controls when evaluating overlap and background (standard IF practice). The catalog paraffin-section caption establishes an IHC example, whereas the HPA nuclear-speckle IF observation does not validate this catalog antibody for IF (caption A30549; HPA subcellular).
How do I distinguish ITPKC signal from chromogenic background?
Run a no-primary section to reveal detection-system background and inspect unstained tissue for pigment before assigning brown deposits to ITPKC (standard IHC practice). Block endogenous peroxidase for a peroxidase-based DAB workflow, then keep chromogen development comparable across sections (standard IHC practice). Check whether staining follows intact cells and plausible nuclear or cytoplasmic compartments rather than folds, edges or damaged tissue (UniProt Q96DU7; standard IHC practice). The selected image reports an immunogen-peptide preabsorption control; use that observation as supporting specificity evidence for its documented conditions (caption A30549). Background in a different block still requires its own controls and morphological review (standard IHC practice).
What should I normalise to when quantifying ITPKC tissue staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then measure nuclear and cytoplasmic staining separately because both compartments are reported for ITPKC (UniProt Q96DU7; HPA tissue IHC; standard IHC practice). For cell-based scoring, report the percentage of positive viable cells or an H-score spanning 0–300, with intensity criteria fixed before review (standard IHC practice). For counted positive cells, express density per mm² of evaluable tissue and exclude necrosis, folds and empty spaces (standard IHC practice). Normalise comparisons to the same eligible cell population or viable tissue area, using matched retrieval, detection and chromogen development (standard IHC practice). Report compartment-specific results alongside representative images (standard IHC practice).
Which staining patterns support genuine ITPKC expression in tissue?
Give greatest weight to reproducible staining in intact cells with nuclear or cytoplasmic distribution, since ITPKC shuttles between those compartments (UniProt Q96DU7). Consider cell identity: HPA reports medium staining in bronchial respiratory epithelial cells but low staining in lung alveolar cells, so adjacent populations need separate assessment (HPA tissue IHC). Treat isolated edge staining, necrotic deposits and signal reproduced on the no-primary control as artefact candidates (standard IHC practice). An endogenous-peroxidase control helps assess misleading DAB signal in a peroxidase workflow (standard IHC practice). Peptide preabsorption reduced staining in the documented image, while HPA reports only medium staining–RNA consistency; interpret a new specimen with its own controls (caption A30549; HPA tissue IHC).
Boster reagents
Best ITPKC / Inositol-trisphosphate 3-kinase C IHC Antibodies
Two anti-ITPKC antibodies list human, mouse and rat reactivity (catalog: A07845-1, A30549). A30549 includes a human paraffin-section IHC image; IF/ICC is listed without an image (A30549 catalog and image caption).
Real IHC data
Anti-IP3KC ITPKC Antibody
Cat # A30549
A07845-1 lists IHC and human, mouse and rat reactivity; no IHC image is supplied (A07845-1 catalog). A30549 lists IHC, IF and ICC with human, mouse and rat reactivity, and its IHC image shows human lung cancer paraffin sections (A30549 catalog and image caption).
Which to pick: For tissue IHC, choose A30549 for its documented human paraffin-section image with Tris-EDTA, pH 8.0 retrieval; the fixative is unreported (A30549 image caption). For IF/ICC, choose A30549 because both applications are listed, although no IF image is supplied (A30549 catalog). Both SKUs list mouse and rat reactivity, but the supplied IHC image documents human tissue only; validate staining in mouse or rat sections for the intended experiment (catalog reactivity; A30549 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96DU7 (IP3KC_HUMAN, Inositol-trisphosphate 3-kinase C).
- Human Protein Atlas. ITPKC tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ITPKC subcellular location (ICC-IF): Localized to the nuclear speckles..
- Human Protein Atlas. ITPKC antibody validation summary (2 antibodies).
- Mendelian Randomization and Transcriptome Analysis Identify Ischemic Stroke Biomarkers With Putative Relevance to Cerebrospinal Fluid. BioMed research international 2026 — PMC13343476.
- PubMed PMID:11085927 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:12747803 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ITPKC (Inositol-trisphosphate 3-kinase C). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30549). Validated IHC antibody: A30549 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 44 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human lung cancer tissue; fixative not specified (datasheet A30549); 2. Fixation: Image fixative and duration unreported (datasheet A30549); 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: Tris-EDTA pH 8.0 (datasheet A30549); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-ITPKC, 1:100 - 1:300 (datasheet A30549); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: High pancreatic expression, but low exocrine-cell staining (UniProt; HPA tissue IHC); Isoform / epitope: No annotated isoforms; one chain with no transmembrane segment (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q96DU7, HPA tissue IHC · Approved, HPA ICC-IF · nuclear speckles., Datasheet A30549, PMC13343476. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ITPKC/immunohistochemistry-ihc-guide.