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PI4KB / Phosphatidylinositol 4-kinase beta · IHC design guide
Design Immunohistochemistry for PI4KB
Plan PI4KB paraffin IHC around cytoplasmic staining in most tissues (HPA tissue IHC). The catalog antibody has an IHC dilution of 2–5 μg/ml (datasheet A04249-2); compare staining across relevant cell types and tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PI4KB 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Adipocytes and glandular cells: cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04249-2) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Cervix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Ovarian follicle cells undetected despite high ovarian expression (HPA tissue IHC; UniProt) | |
| Regulation | No intensity regulator specified (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage unknown (UniProt) |
Section 1
Recommended PI4KB IHC & IF Protocols
The catalog antibody protocol specifies EDTA pH 8.0 heat retrieval (datasheet A04249-2). One published PI4KB IHC protocol qualifies for a parameter table (PMC12134136).
| Sample | Paraffin-embedded human skeletal muscle tissue; fixative not specified (datasheet A04249-2) |
| Fixation | Image fixative and duration unreported (datasheet A04249-2); 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 A04249-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04249-2) |
| Primary antibody | Rabbit anti-PI4KB, 2-5 μg/ml (datasheet A04249-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04249-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04249-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PI4KB-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A04249-2); the published protocol does not state a retrieval method (PMC12134136).
Section 2
What Is the Expected PI4KB Staining Pattern?
PI4KB should appear mainly cytoplasmic in tissue sections, with a perinuclear or Golgi emphasis possible because it associates with endomembranes despite having no transmembrane segment (HPA tissue IHC; UniProt Q9UBF8 topology and subcellular location). Look for staining in the specific cell populations documented by HPA, such as adipocytes, neurons and glandular cells. The tissue IHC profile is Approved, with medium consistency between staining and RNA data (HPA tissue IHC).
| Cytoplasmic staining in adipocytes, neuronal cells or glandular cells; sometimes concentrated near the nucleus. | This fits the reported tissue profile and PI4KB's Golgi and perinuclear localisation (HPA tissue IHC; UniProt Q9UBF8 subcellular location). HPA lists medium staining in adipose adipocytes, cortical neurons and duodenal glandular cells; compare the matching cell population, not the entire section (HPA tissue IHC). |
| Predominantly nuclear staining, or a crisp plasma membrane rim without appreciable cytoplasmic staining. | These compartments do not match the reported cytoplasmic tissue profile or supported Golgi localisation (HPA tissue IHC; HPA subcellular ICC-IF). Treat the pattern as suspect; review morphology and controls before scoring it as PI4KB. Membrane association alone does not predict a cell surface rim (UniProt Q9UBF8 subcellular location). |
| Strong staining in a cell population HPA lists as not detected, such as esophageal squamous cells or endometrial stromal cells. | Possible explanations include antibody cross-reactivity or endogenous detection activity; staining alone cannot distinguish them (general IHC practice). HPA's finding is specific to the named cells and should not be extended to every cell in that tissue (HPA tissue IHC). |
| Weak, widespread colour across cells and empty spaces, obscuring intracellular boundaries. | This is more consistent with diffuse assay background than the reported cytoplasmic cellular pattern (HPA tissue IHC; general IHC practice). Examine a no-primary control and the distribution of signal relative to cell outlines before interpreting intensity (general IHC practice). |
| No visible signal in a documented medium-staining cell population, such as adipose adipocytes or cerebellar Purkinje cells. | The result does not reproduce that HPA observation, but one negative section does not establish absent protein (HPA tissue IHC). Verify that the intended cells are present, then check assay controls and detection performance (general IHC practice). HPA rates tissue IHC consistency with RNA as medium (HPA tissue IHC). |
💡Expected PI4KB appearanceCall a section positive when identifiable HPA-listed cells show cytoplasmic staining, potentially stronger around the Golgi or nucleus, with medium staining plausible in the listed examples; isolated nuclear staining, a surface rim or diffuse colour beyond cells is a suspect result (HPA tissue IHC; HPA subcellular ICC-IF; UniProt Q9UBF8 subcellular location; general IHC practice).
| Cell population and tissue choice | HPA reports medium staining in selected adipocytes, neurons and glandular cells, low staining in skeletal myocytes and kidney tubule cells, and no detection in specific other populations (HPA tissue IHC). Score the identified cells against their own reference level; a low-staining population is a weak positive comparator. |
| Compartment and topology | UniProt places PI4KB at the Golgi, rough ER and mitochondrial outer membrane and in the perinuclear cytoplasm, while reporting no transmembrane segment (UniProt Q9UBF8 topology and subcellular location). HPA describes tissue staining broadly as cytoplasmic and supports Golgi localisation by ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). |
| Differences between source summaries | UniProt describes broad expression with high levels in ovary, yet HPA reports no detection in ovarian follicle cells (UniProt Q9UBF8 tissue specificity; HPA tissue IHC). These statements cover different kinds of evidence and cellular scope; do not convert whole-tissue expression into an expected positive follicle-cell stain. |
| Antibody evidence | The three listed antibodies have Approved IHC status; only CAB022071 also has Supported ICC status (HPA antibody validation). Approved tissue IHC carries medium staining–RNA consistency in this record (HPA tissue IHC). Interpret unusual staining with controls rather than treating the status as proof that every stained cell is specific. |
| Isoforms and epitope coverage | UniProt lists three PI4KB isoforms and a processed chain spanning residues 2–816 (UniProt Q9UBF8 isoforms and processing). No antibody epitope or isoform coverage is supplied, so this record cannot predict whether the catalog antibody distinguishes isoforms or whether a particular tissue result reflects epitope selection. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA medium-staining cells remain blank. | The observed result conflicts with the selected HPA reference population; the source does not establish a PI4KB-specific cause (HPA tissue IHC). | Confirm the reference cells are in the section and review positive, no-primary and detection controls before changing general IHC conditions (general IHC practice). |
| Only nuclei or cell borders stain. | The pattern conflicts with cytoplasmic tissue staining and supported Golgi localisation (HPA tissue IHC; HPA subcellular ICC-IF). | Recheck cell boundaries and counterstain, then compare with a no-primary control and an independently validated antibody where available (general IHC practice). |
| A population listed as not detected stains strongly. | Cross-reactivity or endogenous detection activity is possible; the HPA call applies to the specified cells (HPA tissue IHC; general IHC practice). | Identify the stained cell type precisely and compare no-primary and detection controls before assigning PI4KB positivity (general IHC practice). |
| Colour is diffuse across tissue and blank spaces. | Noncellular or widespread background prevents a reliable match to HPA's cytoplasmic pattern (HPA tissue IHC; general IHC practice). | Inspect a no-primary control and reassess blocking, washes and chromogen development as general IHC workflow checks (general IHC practice). |
| Ovarian follicle cells are negative despite UniProt's high ovarian expression summary. | HPA specifically lists follicle cells as not detected; UniProt's tissue-wide statement does not specify those cells (HPA tissue IHC; UniProt Q9UBF8 tissue specificity). | Report the cell-specific negative result and use an HPA-listed medium-staining population for a positive comparator (HPA tissue IHC). |
| Q: Should IF/ICC show the same diffuse cytoplasmic appearance as IHC? | A: HPA supports Golgi localisation in ICC-IF, while its tissue IHC summary describes cytoplasmic staining in most tissues (HPA subcellular ICC-IF; HPA tissue IHC). | For IF/ICC, assess whether signal concentrates at the Golgi; use the separate IF/ICC guide for assay setup (HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing PI4KB staining
A04249-2 · IHCIHC analysis of PI4KB using anti-PI4KB antibody (A04249-2). PI4KB was detected in a paraffin-embedded section of human skeletal muscle tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000143393-PI4KB/tissue/cerebral+cortex#imid_2012312
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000143393-PI4KB/tissue/colon#imid_2012246
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000143393-PI4KB/tissue/liver#imid_2012247
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000143393-PI4KB/tissue/kidney#imid_2012248
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022071; green = PI4KB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022071; green = PI4KB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PI4KB IHC & IF
🧪Run adipose tissue first: adipocytes should stain (HPA: Medium in adipocytes). Use cervix glandular cells as a negative comparator (HPA: Not detected); unstained cells on the adipose slide can show background, but their PI4KB-negative status is unvalidated.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Cervix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PI4KB in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched nonimmune rabbit IgG control (hero caption: rabbit primary antibody); use PI4KB knockout material, if available, or validated immunizing-peptide competition as a biological specificity control. For chromogenic detection, quench endogenous peroxidase and assess background around adipocyte lipid vacuoles (HPA: adipose tissue positive; hero caption: peroxidase/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A04249-2 paraffin-section caption does not state a fixative (hero caption: fixative unreported). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; HPA ICC-IF images support a Golgi readout in A-431, U-251MG and U2OS, while the supplied evidence does not establish that IF or frozen sections are easier (hero caption: EDTA retrieval; HPA: supported Golgi localization). In adipose sections, check whether staining follows adipocyte lipid-vacuole edges rather than a specific cellular pattern (HPA: Medium in adipocytes).
HPA tissue IHC evidence for PI4KB
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 → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | 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 → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PI4KB IHC Tips
Troubleshoot PI4KB staining by checking retrieval, cellular distribution and controls against the catalog antibody’s tissue-IHC example and independent expression evidence.
Which retrieval conditions should I try first for PI4KB in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A04249-2). The catalog antibody stained a paraffin section of human skeletal muscle after this retrieval, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A04249-2). If staining is weak, compare retrieval heating and cooling conditions on adjacent sections while holding antibody concentration and DAB development constant; include a no-primary control to reveal detection background. Score cellular staining against morphology, because HPA reports only low staining in skeletal-muscle myocytes even though the catalog image shows detectable PI4KB (HPA tissue IHC; datasheet A04249-2).
Could fixation explain weak or uneven PI4KB staining?
The catalog example identifies a paraffin section but does not state its fixative, so PI4KB sensitivity to fixation in this assay is unknown (datasheet A04249-2). Record the fixative, fixation duration, tissue thickness and processing history for each specimen before comparing staining across blocks. On serial sections, keep EDTA retrieval at pH 8.0, primary antibody at 2 μg/ml and DAB development consistent with the catalog starting conditions (datasheet A04249-2). If a batch looks weak, compare a previously responsive block in the same run and inspect tissue preservation; neither HPA expression patterns nor PI4KB membrane association establishes a fixation effect (HPA tissue IHC; UniProt Q9UBF8 subcellular location).
What cellular staining pattern is plausible for PI4KB?
Assess PI4KB chiefly as cytoplasmic staining with possible perinuclear or Golgi enrichment: HPA reports cytoplasmic expression in most tissues and supported Golgi localisation by ICC/IF (HPA tissue IHC; HPA subcellular). UniProt also places PI4KB at the Golgi, rough endoplasmic reticulum and mitochondrial outer membrane, and describes recruitment to the Golgi by ARF and ACBD3 (UniProt Q9UBF8 subcellular location). Chromogenic DAB cannot by itself resolve those closely spaced membrane compartments, so score a reproducible cellular pattern without assigning individual brown puncta to an organelle. If staining is predominantly nuclear or confined to section edges, review morphology and no-primary controls before calling it PI4KB-specific.
Can this antibody distinguish PI4KB isoforms or modified epitopes in sections?
PI4KB has 3 listed isoforms, but the supplied catalog caption gives no immunogen sequence or mapped epitope (UniProt Q9UBF8 isoforms; datasheet A04249-2). Do not assign an IHC difference to a particular isoform without checking whether the antibody epitope is shared among the relevant sequences. PI4KB includes a PIK helical region at residues 29–242, a catalytic region at 535–801, and annotated phosphorylation sites, including serine 258 and threonine 263 (UniProt Q9UBF8 domains; modified residues). These annotations identify questions for epitope mapping; they do not show that retrieval exposes a particular site or that staining measures phosphorylation.
How should I adapt PI4KB assessment for multiplex IF?
Treat IF as a separate optimisation: the supplied catalog example establishes chromogenic staining in a paraffin section, while HPA reports supported Golgi localisation by ICC/IF (datasheet A04249-2; HPA subcellular). Pair PI4KB with a marker for the cell population under study, then inspect whether the PI4KB signal falls within those marker-positive cells rather than relying on tissue-wide fluorescence. Choose fluorophores and image settings after checking an unstained section for tissue autofluorescence, and include single-stain controls to assess spectral bleed-through. PI4KB lacks a transmembrane segment but associates with intracellular membranes, so titrate permeabilisation to make the antibody’s unknown epitope accessible while preserving Golgi morphology (UniProt Q9UBF8 topology; subcellular location).
How can I reduce diffuse brown background without losing PI4KB signal?
Run no-primary and detection-only controls alongside a section stained with the catalog starting conditions: 10% goat serum block, 2 μg/ml primary antibody overnight at 4°C, and peroxidase-based DAB detection (datasheet A04249-2). Include an appropriate endogenous-peroxidase block and limit DAB development if control sections become brown; these are general chromogenic IHC controls, not PI4KB-specific validation. Compare a primary-antibody titration on adjacent sections while keeping retrieval at EDTA pH 8.0 (datasheet A04249-2). Distinguish diffuse reagent staining from cellular cytoplasmic signal using morphology and controls, since HPA describes PI4KB staining mainly as cytoplasmic across tissues (HPA tissue IHC).
What is a defensible way to quantify PI4KB IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, because HPA describes broad cytoplasmic PI4KB expression but different staining levels among cell populations (HPA tissue IHC). For comparable sections, report the percentage of positive eligible cells and an intensity-weighted H-score from 0–300; alternatively, report positive-cell density per mm² when cell counts vary. Normalise to eligible cells or viable tissue area, and keep retrieval, primary-antibody exposure and DAB development consistent with the chosen assay conditions. Record cytoplasmic and perinuclear staining separately when morphology allows, and exclude folds, edges and necrotic areas using rules set before reviewing group labels.
How do I distinguish credible PI4KB staining from section artefact?
A credible result has cellular cytoplasmic staining, potentially enriched near the Golgi or nucleus, consistent with HPA’s tissue profile and supported Golgi localisation (HPA tissue IHC; HPA subcellular). Check which cells are positive: HPA reports medium staining in adipocytes and several glandular or neuronal populations, but low staining in skeletal-muscle myocytes and no detected staining in several listed cell populations (HPA tissue IHC). Treat nuclear-only deposits, staining limited to cut edges or necrosis, and brown signal in no-primary controls as reasons to investigate artefact. Check endogenous-peroxidase blocking and DAB development when control tissue browns, and avoid interpreting staining intensity alone as PI4KB enzymatic activity.
Boster reagents
Best PI4KB / Phosphatidylinositol 4-kinase beta IHC Antibodies
A04249-2 has real IHC data from human paraffin-embedded skeletal muscle (image caption: A04249-2). Its listed reactivity covers human, mouse and rat (catalog: A04249-2 reactivity).
Real IHC data
Anti-PI4KB Antibody ®
Cat # A04249-2
A04249-2 will render with its own IHC figure from human paraffin-embedded skeletal muscle (image caption: A04249-2). The catalog lists IHC as an application and human, mouse and rat reactivity; the figure documents human tissue only (catalog: A04249-2 applications/reactivity; image caption: A04249-2).
Which to pick: Choose A04249-2 for paraffin-section tissue IHC: its caption documents EDTA retrieval at pH 8.0, 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and HRP/DAB detection in human skeletal muscle (image caption: A04249-2). No IF/ICC application or IF figure is listed, so this payload supports no IF/ICC-validated choice (catalog: A04249-2 applications/IF images). For mouse or rat, A04249-2 has listed reactivity but only a human IHC figure; its host is rabbit, clonality is unreported, and the paraffin-section fixative is unreported (catalog: A04249-2 reactivity/host/clone; image caption: A04249-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UBF8 (PI4KB_HUMAN, Phosphatidylinositol 4-kinase beta).
- Human Protein Atlas. PI4KB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PI4KB subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. PI4KB antibody validation summary (3 antibodies).
- circSLC6A6 Sponges miR-497-5p to Promote Endometrial Cancer Progression via the PI4KB/Hedgehog Axis. Journal of immunology research 2021 — PMC8245255.
- Oncogenic RAS induces a distinctive form of non-canonical autophagy mediated by the P38-ULK1-PI4KB axis. Cell research 2025 — PMC12134136.
- PubMed PMID:9405935 — UniProt-cited evidence.
- PubMed PMID:9405938 — UniProt-cited evidence.
- PubMed PMID:9020160 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PI4KB (Phosphatidylinositol 4-kinase beta). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04249-2). Validated IHC antibody: A04249-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Cervix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human skeletal muscle tissue; fixative not specified (datasheet A04249-2); 2. Fixation: Image fixative and duration unreported (datasheet A04249-2); 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 A04249-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04249-2); 8. Primary antibody: Rabbit anti-PI4KB, 2-5 μg/ml (datasheet A04249-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04249-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Ovarian follicle cells undetected despite high ovarian expression (HPA tissue IHC; UniProt); Isoform / epitope: 3 isoforms; epitope coverage unknown (UniProt); Regulation: No intensity regulator specified (UniProt). Evidence: UniProt Q9UBF8, HPA tissue IHC · Approved, HPA ICC-IF · Golgi apparatus., Datasheet A04249-2, PMC12134136. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PI4KB/immunohistochemistry-ihc-guide.