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PIP5K1A / Phosphatidylinositol 4-phosphate 5-kinase type-1 alpha · IHC design guide
Design Immunohistochemistry for PIP5K1A
Plan paraffin-section IHC for PIP5K1A using its cytoplasmic tissue staining profile (HPA tissue IHC). This guide pairs tissue controls with the catalog antibody’s validated staining conditions (datasheet A07416-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PIP5K1A 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07416-3) | |
| Positive control | Appendix+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 consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Heart expression contrasts with absent cardiomyocyte staining (UniProt; HPA tissue IHC) | |
| Regulation | Calcium-linked membrane recruitment (UniProt) | |
| Isoform / epitope | 4 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended PIP5K1A IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published PIP5K1A tissue microarray protocol (PMC13193874).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A07416-3) |
| Fixation | Image fixative and duration unreported (datasheet A07416-3); 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 A07416-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07416-3) |
| Primary antibody | Rabbit anti-PIP5K1A, 2-5 μg/ml (datasheet A07416-3) |
| Primary incubation | Overnight at 4 °C (datasheet A07416-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07416-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PIP5K1A-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A07416-3).
Section 2
What Is the Expected PIP5K1A Staining Pattern?
PIP5K1A should show mainly cytoplasmic staining in tissue IHC, with membrane-associated signal possible in suitable cells (HPA tissue IHC; UniProt Q99755 localization). Strong examples include germinal center cells in lymph node and tonsil, lymphoid tissue in appendix, and cells in testicular seminiferous ducts (HPA: High). The protein has no transmembrane segment (UniProt Q99755 topology). HPA rates the tissue staining Approved, with medium consistency against RNA expression (HPA tissue IHC).
| Clear cytoplasmic signal in germinal center cells, appendix lymphoid tissue, or cells in seminiferous ducts; nearby structures remain distinguishable. | This matches the strongest supplied tissue observations (HPA: High in these cell populations). Compare staining within each tissue compartment; HPA's Approved rating and medium RNA consistency support a plausible pattern but do not make every positive cell specific (HPA tissue IHC). |
| A membrane rim accompanies cytoplasmic staining, or a limited nuclear component appears in otherwise plausible cells. | Membrane and nuclear localization are biologically compatible with PIP5K1A (UniProt Q99755 localization). HPA ICC-IF supports plasma membrane and cytosol, with additional nucleoplasm (HPA subcellular). Tissue IHC is summarized as cytoplasmic, so use the tissue pattern and controls before interpreting a minor second compartment (HPA tissue IHC). |
| Staining is confined to a compartment unrelated to the supplied localizations, with no credible cytoplasmic or membrane-associated signal. | Treat an isolated, incompatible pattern as a possible artefact and review controls and tissue morphology (UniProt Q99755 localization; standard IHC practice). Nuclear signal alone needs care: nuclear localization is reported, including speckles, although HPA's tissue IHC profile is cytoplasmic (UniProt Q99755 localization; HPA tissue IHC). |
| An unexpected cell population stains strongly while the expected positive population is weak or absent. | Consider cross-reactivity or, for enzyme-based detection, endogenous activity (standard IHC practice). Cell identity matters: HPA reports lung macrophages as Low and heart cardiomyocytes as Not detected, despite broader tissue-level expression statements in UniProt (HPA tissue IHC; UniProt Q99755 tissue specificity). |
| Color spreads across cells, stroma, or section edges without a discernible intracellular pattern, or a known-positive cell population has no signal. | Diffuse staining limits localization and may reflect detection background; absent staining requires a check of the antibody and IHC workflow (standard IHC practice). Use an HPA High population as a reference, then assess morphology and controls before calling the specimen negative (HPA tissue IHC; standard IHC practice). |
💡Expected PIP5K1A appearanceCall a positive result when cytoplasmic staining is clearly resolved in an HPA High population, such as germinal center cells, with possible membrane-associated signal; broad color unrelated to cell boundaries is suspect (HPA tissue IHC; UniProt Q99755 localization; standard IHC practice).
| Tissue and cell selection (HPA tissue IHC) | HPA reports High staining in lymph node and tonsil germinal center cells, appendix lymphoid tissue, and cells in testicular seminiferous ducts. It reports Not detected in adipocytes and heart cardiomyocytes, among other listed populations. Choose comparisons by cell type within the section (HPA tissue IHC). |
| Strength of tissue evidence (HPA tissue IHC) | The tissue profile is Approved, with medium consistency between antibody staining and RNA expression; HPA lists low RNA tissue specificity. These ratings support pattern review but leave individual unusual staining results to be checked against controls and morphology (HPA tissue IHC; standard IHC practice). |
| Location and topology (UniProt Q99755; HPA subcellular) | PIP5K1A is reported at the cell membrane, in cytoplasm, and in the nucleus, including speckles; it has no transmembrane segment (UniProt Q99755 localization and topology). HPA ICC-IF supports plasma membrane and cytosol, with additional nucleoplasm; its tissue IHC summary emphasizes cytoplasm (HPA subcellular; HPA tissue IHC). |
| Protein forms and antibody scope (UniProt Q99755) | UniProt lists four isoforms, a single chain spanning residues 1–562, no signal peptide or propeptide, and no listed glycosylation sites (UniProt Q99755). The supplied record does not locate this antibody's epitope, so these features cannot establish which isoforms it recognizes or predict retrieval behavior. |
| IF/ICC question: should its image match paraffin IHC? (HPA subcellular) | IF/ICC may resolve plasma membrane, cytosol, and additional nucleoplasm in the imaged cell lines A-431, U-251MG, and U2OS (HPA subcellular). Paraffin tissue IHC has a cytoplasmic summary and cell-specific staining levels; compare each application with its own evidence (HPA tissue IHC; HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in an HPA High reference population. | The staining run may have failed, or the chosen section may lack an intact, identifiable reference population (HPA tissue IHC; standard IHC practice). | Confirm the expected cells on the counterstained section and review the catalog antibody's IHC-P conditions, retrieval, dilution, and detection controls before interpreting other tissue as negative (standard IHC practice). |
| A test population is negative, but the HPA High reference stains as expected. | The test population may genuinely show little or no detectable staining; HPA lists several cell populations as Not detected or Low (HPA tissue IHC). | Score the named cell type rather than the organ as a whole, preserve the successful reference result, and report the observed localization and intensity (HPA tissue IHC; standard IHC practice). |
| Only nuclei stain strongly across many unrelated cell types. | Nuclear PIP5K1A is possible, but widespread nuclear-only tissue staining differs from HPA's cytoplasmic IHC summary (UniProt Q99755 localization; HPA tissue IHC). | Check whether expected cytoplasmic staining appears in an HPA High reference and compare negative controls before assigning the nuclear signal to PIP5K1A (HPA tissue IHC; standard IHC practice). |
| An HPA Not detected cell population shows prominent signal. | The observation conflicts with that cell-specific HPA entry; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; standard IHC practice). | Verify the cell identity and morphology, then examine a negative reagent control; if enzyme-based detection is used, check its endogenous-activity control (standard IHC practice). |
| Background obscures cell borders and subcellular localization. | Non-specific binding or detection background can make staining uninterpretable (standard IHC practice). | Review blocking, washing, antibody dilution, and detection controls under the chosen IHC workflow; reassess only when intracellular staining can be separated from background (standard IHC practice). |
| Heart tissue appears positive overall, but cardiomyocytes do not stain. | UniProt reports high expression in heart, while HPA specifically scores cardiomyocytes as Not detected by tissue IHC; the statements measure different scopes (UniProt Q99755 tissue specificity; HPA tissue IHC). | Identify which heart cell population carries the signal and report that observation without assigning it to cardiomyocytes or overriding the cell-specific HPA result (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing PIP5K1A staining
A07416-3 · IHCIHC analysis of PIP5K1A using anti-PIP5K1A antibody (A07416-3). PIP5K1A was detected in a paraffin-embedded section of mouse brain tissue. H
A07416-3 · IHCIHC analysis of PIP5K1A using anti-PIP5K1A antibody (A07416-3). PIP5K1A was detected in a paraffin-embedded section of rat brain tissue. Hea
A07416-3 · IFIF analysis of PIP5K1A using anti-PIP5K1A antibody (A07416-3) and anti-Beta Tubulin antibody (M01857-3). PIP5K1A was detected in an immunocy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000143398-PIP5K1A/tissue/cerebral+cortex#imid_7983935
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000143398-PIP5K1A/tissue/colon#imid_7983869
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000143398-PIP5K1A/tissue/liver#imid_7983870
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000143398-PIP5K1A/tissue/kidney#imid_7983879
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029366; green = PIP5K1A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029366; green = PIP5K1A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PIP5K1A IHC & IF
🧪Run lymph node first and look for staining in germinal center cells (HPA: High in germinal center cells). Use adipose tissue as the negative tissue, with adipocytes near background (HPA: Not detected in adipocytes); on the lymph node slide, use cells outside germinal centers as internal background comparators, without assuming they are PIP5K1A-negative.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PIP5K1A in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, then assess specificity with PIP5K1A knockout tissue or peptide competition if the immunizing peptide is available (standard IHC practice). For HRP/DAB detection in lymph node, quench endogenous peroxidase and check for residual background (standard IHC practice; selected-SKU caption: HRP/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The reported mouse brain IHC example uses heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval as necessary for lymph node (selected-SKU caption: heat retrieval in EDTA, pH 8.0). Frozen sections or IF cannot be judged easier from these data; for separate ICC-IF assessment, plasma membrane and cytosol are supported locations, with nucleoplasm an additional location (HPA subcellular: supported locations).
HPA tissue IHC evidence for PIP5K1A
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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PIP5K1A IHC Tips
Use the catalog antibody’s paraffin-section result as the starting point, then assess staining by compartment and cell type (datasheet A07416-3; UniProt Q99755; HPA tissue IHC).
What retrieval should I try first for PIP5K1A in paraffin sections?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A07416-3). The selected mouse-brain image used that retrieval before overnight incubation with 2 μg/ml catalog antibody at 4°C (datasheet A07416-3). If staining is weak, compare retrieval times on adjacent sections while holding antibody concentration and chromogenic development constant; excessive heating can damage morphology (standard IHC practice). Include a section processed without primary antibody to distinguish retrieval-related background from antibody staining, and assess signal in intact cells rather than relying on tissue edges (standard IHC practice).
How should I troubleshoot fixation when PIP5K1A staining is weak?
The selected paraffin-section caption does not report its fixative, so target-specific fixation sensitivity is unknown (datasheet A07416-3). Record the fixative and fixation duration for each specimen, and compare consistently processed sections before attributing a weak result to PIP5K1A abundance (standard IHC practice). Preserve the documented starting conditions—EDTA at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C—when evaluating a fixation change (datasheet A07416-3). Check morphology, a no-primary control, and staining in the same cell population across conditions; differences alone cannot identify a PIP5K1A-specific fixation effect (standard IHC practice).
Where should convincing PIP5K1A staining appear in an IHC section?
Assess cytoplasmic staining first: the tissue survey reports cytoplasmic expression in most tissues (HPA tissue IHC). Membrane-associated and nuclear signal can also be biologically plausible because PIP5K1A is annotated at the cell membrane, cytoplasm, nucleus, ruffles, lamellipodia, and nuclear speckles (UniProt Q99755 subcellular location). Its lack of a transmembrane segment means a membrane-enriched pattern should not be interpreted as evidence of membrane-spanning topology (UniProt Q99755 topology). Compare compartments within intact cells at the same DAB development time, using the no-primary section to identify staining unrelated to primary-antibody binding (standard IHC practice).
Can this IHC pattern distinguish PIP5K1A isoforms or modified epitopes?
Do not assign an isoform from this stain alone: PIP5K1A has 4 annotated isoforms, while the supplied catalog caption does not specify an antibody epitope or isoform coverage (UniProt Q99755 isoforms; datasheet A07416-3). The canonical protein contains a PIPK domain at residues 81–449 and an annotated phosphoserine at residue 486 (UniProt Q99755 domains and modified residues). Neither a strong nor a weak DAB signal establishes phosphorylation state without a validated modification-specific reagent (standard IHC interpretation). For an isoform-specific claim, first establish the immunogen sequence and its presence in each isoform, then validate specificity independently (standard antibody-validation practice).
How can IF help check a PIP5K1A IHC localisation result?
Use IF as a complementary localisation check, with separate antibody validation for that application; the selected catalog result documents chromogenic IHC in paraffin-embedded mouse brain (datasheet A07416-3). Multiplex PIP5K1A with a validated marker for the cell population being assessed, such as germinal-center cells when examining the reported high-staining lymph-node population (HPA tissue IHC; standard IF practice). Choose a spectrally separated, preferably far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and include single-label controls (standard IF practice). Because PIP5K1A lacks a transmembrane segment and has cytosolic and nuclear locations, optimize permeabilisation to expose intracellular epitopes without losing spatial detail (UniProt Q99755 topology and subcellular location; standard IF practice).
What should I check when PIP5K1A DAB staining looks diffuse?
Run a no-primary section through the same secondary antibody and DAB steps to detect background from the chromogenic workflow (standard IHC practice). The selected caption used 10% goat serum block, a peroxidase-conjugated anti-rabbit secondary for 30 minutes at 37°C, and DAB development (datasheet A07416-3). Check endogenous peroxidase activity with an appropriate block, and compare wash stringency and DAB development on adjacent sections (standard IHC practice). Diffuse cytoplasmic signal is not automatically background, since the tissue survey describes cytoplasmic PIP5K1A expression; judge it alongside cellular morphology, no-primary staining, and compartment boundaries (HPA tissue IHC; standard IHC interpretation).
How should I quantify PIP5K1A staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since the tissue survey reports cytoplasmic expression in most tissues and cell-type differences within organs (HPA tissue IHC). For cellular staining, report the percentage of positive cells and an intensity-weighted H-score on a 0–300 scale; for discrete positive populations, density per mm² of viable tissue is another option (standard IHC scoring practice). Normalize counts to the number of eligible cells or measured viable area, and use identical thresholds, retrieval, antibody incubation, and DAB development across samples (standard IHC practice). Score tissue edges and damaged regions separately or exclude them by a preset rule (standard IHC practice).
How can I distinguish a true PIP5K1A-positive cell from artefact?
A convincing result follows intact cell boundaries and a plausible compartment: cytoplasm is common in tissue IHC, while membrane-associated and nuclear localisation are also annotated (HPA tissue IHC; UniProt Q99755 subcellular location). Check that the stained cell type matches the claim; germinal-center cells show high staining in lymph node, whereas adipocytes are reported as not detected (HPA tissue IHC). Discount staining concentrated at section edges, necrotic areas, or tissue folds, and compare with a no-primary control for endogenous enzyme or secondary-antibody signal (standard IHC practice). Treat a discrepant pattern cautiously because the tissue atlas rates antibody staining versus RNA expression as having medium consistency (HPA tissue IHC).
Boster reagents
Best PIP5K1A / Phosphatidylinositol 4-phosphate 5-kinase type-1 alpha IHC Antibodies
The IHC-validated antibody has paraffin-section images from human liver and liver cancer, mouse brain, and rat brain, plus an IF image from HeLa cells (catalog image captions).
Real IHC data
Anti-PIP5K1A Antibody ®
Cat # A07416-3
A07416-3 has IHC images from paraffin sections of mouse and rat brain and human liver and liver cancer (A07416-3 IHC captions). A07416-3 also has an IF image from HeLa cells and lists IHC, ICC, and IF among its applications (A07416-3 IF caption; catalog applications).
Which to pick: Choose A07416-3 for tissue IHC on paraffin sections; its captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (A07416-3 IHC captions). Choose the same SKU for IF/ICC based on its HeLa cell IF image and listed ICC/IF applications (A07416-3 IF caption; catalog applications). For cross-species work, it lists human, mouse, and rat reactivity and has IHC images from all three species (A07416-3 catalog reactivity; IHC captions); its clone and the tissue fixative are unreported (A07416-3 catalog record; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99755 (PI51A_HUMAN, Phosphatidylinositol 4-phosphate 5-kinase type-1 alpha).
- Human Protein Atlas. PIP5K1A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PIP5K1A subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. PIP5K1A antibody validation summary (1 antibodies).
- Targeting PIP5K ameliorates hepatic cancer by inhibiting PI3K/AKT and the autophagy mechanism and enhancing ROS-mediated apoptosis. Frontiers in pharmacology 2026 — PMC13193874.
- PIP5K1A Suppresses Ferroptosis and Induces Sorafenib Resistance by Stabilizing NRF2 in Hepatocellular Carcinoma. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2025 — PMC12376573.
- Circ_PIP5K1A regulates cisplatin resistance and malignant progression in non-small cell lung cancer cells and xenograft murine model via depending on miR-493-5p/ROCK1 axis. Respiratory research 2021 — PMC8449484.
- PubMed PMID:8955136 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PIP5K1A (Phosphatidylinositol 4-phosphate 5-kinase type-1 alpha). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07416-3). Validated IHC antibody: A07416-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A07416-3); 2. Fixation: Image fixative and duration unreported (datasheet A07416-3); 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 A07416-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07416-3); 8. Primary antibody: Rabbit anti-PIP5K1A, 2-5 μg/ml (datasheet A07416-3); 9. Primary incubation: Overnight at 4 °C (datasheet A07416-3). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Heart expression contrasts with absent cardiomyocyte staining (UniProt; HPA tissue IHC); Isoform / epitope: 4 isoforms; check epitope coverage (UniProt); Regulation: Calcium-linked membrane recruitment (UniProt). Evidence: UniProt Q99755, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A07416-3, PMC13193874. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PIP5K1A/immunohistochemistry-ihc-guide.