PIP4K2A / Phosphatidylinositol 5-phosphate 4-kinase type-2 alpha · IHC design guide

Design Immunohistochemistry for PIP4K2A

Plan paraffin-section PIP4K2A IHC around the general nuclear pattern, with high staining reported in brain glial cells (HPA tissue IHC). The catalog antibody has a documented human brain IHC-P example at 1:25 with DAB detection (datasheet: A04244-1).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for PIP4K2A (IHC for PIP4K2A): expected localisation General nuclear tissue staining (HPA tissue IHC), antibody A04244, validated IHC image, and IHC protocol steps — Printable **PIP4K2A** IHC protocol sheet — expected localisation General nuclear tissue staining (HPA tissue IHC), antibody A04244, controls and protocol steps. Open the full PIP4K2A IHC guide →

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PIP4K2A 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 nuclear tissue staining (HPA tissue IHC)
	Staining pattern	General nuclear staining across tissues (HPA tissue IHC)
	Antigen retrieval	Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen)
	Positive control ⓘ	Adrenal gland+4 more · see all
	Negative control ⓘ	None in HPA (detected in all 45 tissues); use no-primary + isotype controls

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep 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 A04244)
	Caveat	Staining has medium consistency with RNA expression (HPA tissue IHC)
	Regulation	Brain-enhanced RNA expression (HPA tissue RNA)
	Isoform / epitope	2 isoforms; check epitope coverage for each (UniProt)

Section 1

Recommended PIP4K2A IHC & IF Protocols

The catalog antibody protocol is followed by published PIP4K2A IHC conditions for cutaneous melanoma and glioma (PMC11585527; PMC6504209).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A04244)
Fixation	Image fixative and duration unreported (datasheet A04244); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen)
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-PIP4K2A, 1:50-1:200 (datasheet A04244)
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	PIP4K2A-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control.

💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval); compare citrate pH 6.0 if optimization is needed (PMC11585527).

Section 2

What Is the Expected PIP4K2A Staining Pattern?

PIP4K2A has no transmembrane segment and is reported in the nucleus, cytoplasm, lysosome and at the cell membrane (UniProt P48426 topology and subcellular location). In paraffin-section IHC, expect predominantly nuclear staining in appropriate tissue cells (HPA: general nuclear expression). HPA rates the tissue IHC pattern “Supported,” with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).

What am I looking at on my slide?
Clear nuclear stain in adrenal or appendix glandular cells, bronchial respiratory epithelium, or marrow hematopoietic cells (HPA: High in each listed cell type).	This fits the reported general nuclear IHC pattern (HPA: tissue IHC). Score the relevant cells and their nuclear intensity; visible stain in other compartments does not automatically invalidate the slide because UniProt also reports cytoplasmic and membrane locations (UniProt P48426 subcellular location).
Strong stain restricted to extracellular material or glandular lumina, with nuclei of expected positive cells unstained.	That distribution conflicts with the reported nuclear IHC pattern and intracellular locations (HPA: general nuclear expression; UniProt P48426 subcellular location). Treat it as a possible artefact until a known-positive section and detection controls support a specific signal (general IHC practice).
Strong adipocyte staining in breast or adipose tissue, especially when expected positive cell populations show little signal (HPA: Low in adipocytes).	This is discordant with HPA’s low adipocyte staining, but “Low” does not mean absent (HPA: breast and adipose tissue). Consider cross-reactivity or endogenous detection activity; compare the cell pattern with a no-primary control before assigning a cause (general IHC practice).
Uniform haze across cells, connective tissue and empty spaces obscures nuclear boundaries.	The pattern cannot be scored as the reported general nuclear expression (HPA: tissue IHC). It suggests nonspecific staining or detection background; a no-primary control helps distinguish detection background from staining associated with the primary antibody (general IHC practice).
No visible signal in a section containing adrenal glandular cells or marrow hematopoietic cells (HPA: High in those cell types).	This is an unexpected result for the selected positive tissue, although HPA reports only medium consistency between staining and RNA data (HPA: tissue IHC reliability). Check section quality and the IHC staining workflow before interpreting an experimental sample as PIP4K2A negative (general IHC practice).

💡Expected PIP4K2A appearanceA convincing positive is discernible nuclear chromogen in an HPA high-staining cell population, such as adrenal glandular cells, with interpretable background; strong stain limited to lumina or empty spaces is suspect (HPA: general nuclear expression; HPA: High in adrenal glandular cells; general IHC practice).

How each factor affects the staining
Which tissue and cells provide a positive comparison?	HPA reports High IHC staining in adrenal and appendix glandular cells, bronchial respiratory epithelial cells, marrow hematopoietic cells, and several glial populations (HPA: tissue IHC). Identify the expected cell population on the section before scoring, rather than treating every cell in a positive tissue as positive (general IHC practice).
What can the compartment tell you?	HPA describes a general nuclear tissue IHC pattern, while UniProt also lists cytoplasm, lysosome and cell membrane (HPA: tissue IHC; UniProt P48426 subcellular location). Predominant nuclear stain supports the tissue-IHC expectation; additional cellular stain needs context rather than automatic rejection.
How strong is the validation evidence?	The tissue IHC profile is “Supported,” with medium staining–RNA consistency; the listed IHC antibody HPA068771 is also “Supported” (HPA: tissue IHC reliability; HPA: antibody validation). These ratings support using the reported pattern as a comparison, while leaving room to investigate discordant sections.
Does topology imply a cell-surface-only pattern?	No transmembrane segment is annotated, and UniProt reports both intracellular and cell-membrane locations (UniProt P48426 topology and subcellular location). A surface-only chromogenic pattern would need independent validation before it replaces the reported nuclear tissue-IHC expectation (HPA: general nuclear expression).
Why might IF/ICC look different from tissue IHC?	HPA’s ICC-IF data place PIP4K2A mainly at the Golgi apparatus, with additional plasma-membrane and cytosolic signal; its tissue IHC profile is generally nuclear (HPA: subcellular ICC-IF; HPA: tissue IHC). Compare results within each assay’s evidence rather than requiring identical compartment patterns.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive tissue has no nuclear stain (HPA: High in adrenal glandular or marrow hematopoietic cells).	The tissue result may reflect an unsuccessful IHC run or an unsuitable section; the supplied sources do not establish a PIP4K2A-specific fixation effect (general IHC practice; HPA: tissue IHC).	Confirm the expected cells are present, review section and counterstain quality, and check retrieval, primary-antibody application and detection against the validated IHC workflow (general IHC practice).
Brown chromogen appears broadly, including where no cells are present.	Detection background or nonspecific staining can obscure the expected nuclear pattern (general IHC practice; HPA: general nuclear expression).	Run a no-primary control; if it also stains, check blocking and detection steps, including endogenous enzyme blocking when enzyme-based detection is used (general IHC practice).
The expected cells stain, but nuclei cannot be distinguished from surrounding cytoplasm.	Overdeveloped chromogen or diffuse background may prevent compartment scoring; cytoplasmic PIP4K2A is also reported (general IHC practice; UniProt P48426 subcellular location).	Review chromogen development, dilution and counterstain using the validated IHC workflow; compare with a no-primary control before calling the pattern nuclear or cytoplasmic (general IHC practice).
Only luminal or extracellular material stains while expected positive nuclei remain clear.	This distribution conflicts with HPA’s general nuclear tissue pattern and UniProt’s listed cellular locations (HPA: tissue IHC; UniProt P48426 subcellular location).	Inspect morphology and compare a known-positive section plus a no-primary control before interpreting the luminal stain as PIP4K2A (general IHC practice).
Adipocytes stain as strongly as nearby expected positive cells (HPA: Low in breast and adipose adipocytes).	The intensity is discordant with HPA’s low adipocyte observation; cross-reactivity or detection background remains possible (HPA: tissue IHC; general IHC practice).	Compare matched no-primary and positive-tissue sections, then report adipocyte staining as uncertain if specificity remains unresolved (general IHC practice).
ICC-IF shows Golgi signal, but the paraffin-section IHC result is predominantly nuclear.	Those are the respective HPA assay observations; the supplied evidence does not establish that one pattern should replace the other (HPA: subcellular ICC-IF; HPA: tissue IHC).	Interpret the paraffin section against the nuclear tissue-IHC profile and its “Supported” reliability; use the ICC-IF localization as assay-specific context (HPA: tissue IHC reliability; HPA: subcellular ICC-IF).

Source: UniProt P48426 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing PIP4K2A staining

Immunohistochemistry of paraffin-embedded rat kidney using PIP4K2A antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

A04244 · IHCImmunohistochemistry of paraffin-embedded rat kidney using PIP4K2A antibody at dilution of 1:100 .Perform microwave antigen retrieval with 1

Immunohistochemistry of paraffin-embedded mouse lung using PIP4K2A antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

A04244 · IHCImmunohistochemistry of paraffin-embedded mouse lung using PIP4K2A antibody at dilution of 1:100 .Perform microwave antigen retrieval with 1

A04244 · IFImmunofluorescence analysis of A549 cells using PIP4K2A antibody .

Immunohistochemical analysis of paraffin-embedded H. brain section using PIP4K2A Antibody (C-term). A04244-1 was diluted at 1:25 dilution. A undiluted biotinylated goat polyvalent antibody was used as the secondary, followed by DAB staining.

A04244-1 · IHCImmunohistochemical analysis of paraffin-embedded H. brain section using PIP4K2A Antibody (C-term). A04244-1 was diluted at 1:25 dilution. A

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized Hela (Human Cervical epithelial adenocarcinoma cell line) cells labeling PIP4K2A with A04244-1 at 1/25 dilution, followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody at 1/400 dilution (green). Confocal image showing cytoplasmic staining on Hela cell line. Cytoplasmic actin is detected with Alexa Fluor® 555 conjugated with Phalloidin at 1/100 dilution (red).

A04244-1 · IFImmunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized Hela (Human Cervical epithelial adenocarcinoma cel

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000150867-PIP4K2A/tissue/cerebral+cortex#imid_19210919

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000150867-PIP4K2A/tissue/colon#imid_19210845

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000150867-PIP4K2A/tissue/liver#imid_19210846

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000150867-PIP4K2A/tissue/kidney#imid_19210847

HPA ICC-IF BJ [Human fibroblast] — PIP4K2A (HPA065440)

HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA065440; green = PIP4K2A, blue = nucleus, red = microtubules.

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Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for PIP4K2A IHC & IF

🧪Run bone marrow first and look for staining in hematopoietic cells (HPA: High in bone marrow hematopoietic cells). HPA detects PIP4K2A in all 45 scored tissues, so use no-primary and isotype controls instead of a negative tissue; cells without specific staining on the positive slide should show counterstain only, but no particular cell type is established as target-negative (HPA: no negative tissue rows).

Positive control tissue: Adrenal gland (Glandular cells, HPA High)

Negative control tissue: None in HPA: PIP4K2A is detected in all 45 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 PIP4K2A in BJ [Human fibroblast], SK-MEL-30, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Golgi apparatus (approved) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only slide and a control matched to the primary antibody’s host and isotype for a monoclonal antibody, or normal IgG from the same host for a polyclonal antibody; confirm specificity with PIP4K2A knockout material. For chromogenic bone marrow IHC, block endogenous peroxidase and check the no-primary slide for residual signal (HPA: High in bone marrow hematopoietic cells).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04244 paraffin-section caption does not state a fixative (caption: fixative not stated). That caption uses microwave retrieval in 10 mM PBS, pH 7.2, with antibody at 1:100, but does not establish that retrieval is required (caption: rat kidney IHC). The supplied evidence does not establish whether frozen sections or IF are easier; in bone marrow, endogenous peroxidase can complicate chromogenic interpretation.

HPA tissue IHC evidence for PIP4K2A

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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	High	Protein (IHC)	HPA →
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	High	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	High	Protein (IHC)	HPA →
Caudate	Glial cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
None in HPA: PIP4K2A is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

Section 3

Advanced PIP4K2A IHC Tips

Troubleshoot PIP4K2A chromogenic IHC in paraffin sections by checking retrieval, compartment patterns, controls and scoring before interpreting staining.

How should I retrieve PIP4K2A in paraffin sections when nuclear staining is weak?

Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval rule). Compare a retrieved section with an otherwise matched section processed without heat, and keep detection conditions constant so a change in nuclear signal can be attributed to retrieval (standard IHC practice). If staining remains weak, test microwave retrieval in 10 mM PBS at pH 7.2, the condition reported for paraffin-embedded rat kidney with catalog antibody A04244 (A04244 tissue-IHC caption). Assess both nuclear signal and tissue damage across conditions; nuclear expression is reported in tissue IHC, but retrieval performance can vary with processing (HPA tissue IHC; standard IHC practice).

Could fixation explain weak or uneven PIP4K2A staining?

Target-specific fixation sensitivity is unknown because the supplied PIP4K2A tissue-IHC caption reports a paraffin section without stating its fixative (A04244 tissue-IHC caption). Record the fixative, fixation duration and processing history for each specimen, then compare matched sections with the same retrieval and detection settings (standard IHC practice). Examine whether weak staining follows a processing batch or tissue depth, and inspect morphology before changing antibody concentration (standard IHC practice). If specimens differ in fixation history, avoid treating their staining intensities as directly comparable until a common control shows that processing has preserved the assay response (standard IHC practice).

How should I assess nuclear, cytoplasmic and membrane staining for PIP4K2A?

First identify the stained cell type and score nuclear and nonnuclear compartments separately; general nuclear expression has been reported in tissue IHC (HPA tissue IHC). Cytoplasmic, cell-membrane and lysosomal locations also appear in the protein record, while its topology lists no transmembrane segment (UniProt P48426 subcellular; UniProt P48426 topology). A shift toward membrane staining can be biologically plausible because tyrosine phosphorylation may promote cytosol-to-membrane translocation (UniProt P48426 subcellular). Compare the pattern with a matched control section and intact morphology, and do not call a sharp tissue-edge rim or diffuse staining across all compartments a compartment shift without further validation (standard IHC practice).

Can this antibody distinguish PIP4K2A isoforms or a modified epitope in tissue?

Do not assign an isoform from chromogenic staining alone: the record lists 2 isoforms but supplies no isoform-specific epitope for catalog antibody A04244 (UniProt P48426 isoforms; A04244 tissue-IHC caption). Request the immunogen sequence or mapped epitope and check whether it is shared by both isoforms before making an isoform-level claim (standard IHC practice). The record lists phosphorylation and acetylation sites, including Thr3, Ser14, Lys89 and Lys145, but supplies no evidence that these modifications alter this antibody’s binding (UniProt P48426 modified residues). If specificity matters, compare staining with an independently validated reagent or an appropriate loss-of-target control, using identical tissue processing (standard IHC practice).

How can I check a PIP4K2A tissue-IHC pattern with multiplex immunofluorescence?

Choose a marker for the cell type being examined, then ask whether PIP4K2A signal occurs in those cells and in which compartment; tissue IHC reports high staining in glial cells in cerebral cortex (HPA tissue IHC). Select a fluorophore whose signal can be separated from the specimen’s autofluorescence and include single-stain and no-primary controls when assessing bleed-through or background (standard IF practice). For a cytosolic or nuclear epitope, use controlled permeabilisation, whereas a surface-accessible epitope may require a different approach; the mapped epitope for catalog antibody A04244 is not supplied (standard IF practice; A04244 tissue-IHC caption). Optimise fixation and permeabilisation for the IF assay itself, because the supplied caption documents tissue IHC only (A04244 tissue-IHC caption).

What should I change when PIP4K2A IHC has diffuse brown background?

Check a no-primary section to locate signal arising from the detection system, then compare it with the stained section at the same counterstain and imaging settings (standard chromogenic IHC practice). Apply a peroxidase block before horseradish-peroxidase detection and adjust protein blocking, washes or antibody concentration if nonspecific colour persists (standard chromogenic IHC practice). For catalog antibody A04244, the rat-kidney paraffin caption reports 1:100 with microwave retrieval in 10 mM PBS at pH 7.2; it does not establish an optimal dilution for other tissues (A04244 tissue-IHC caption). Inspect tissue edges and damaged regions separately before interpreting widespread brown deposit as PIP4K2A expression (standard IHC practice).

How should I quantify PIP4K2A staining across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, since tissue IHC reports general nuclear expression and high staining in specified cell types, including bone-marrow hematopoietic cells (HPA tissue IHC). Within a consistent region of interest, record the percentage of positive cells and staining intensity, then calculate an H-score if ordinal intensity categories were defined in advance (standard IHC practice). For spatial comparisons, report positive-cell density per mm² and normalise counts to the evaluated viable tissue area or eligible cell population (standard IHC practice). Keep retrieval, chromogen development, counterstain and image thresholds consistent, and exclude folds, necrosis and edge artefacts by a prespecified rule (standard IHC practice).

How can I distinguish true PIP4K2A staining from artefact?

A credible result should show reproducible staining in intact cells with a compartment pattern that can be assessed against reported general nuclear tissue expression (HPA tissue IHC; standard IHC practice). Check whether the signal belongs to the cells being scored: high staining has been reported in adrenal glandular cells and cerebral-cortex glial cells, while low staining was reported in adipocytes (HPA tissue IHC). Compare serial sections and controls before accepting staining confined to cut edges, necrotic areas or damaged tissue as target signal (standard IHC practice). If brown colour also appears in a no-primary control, investigate endogenous enzyme activity or detection background before drawing a biological conclusion (standard chromogenic IHC practice).

Boster reagents

Best PIP4K2A / Phosphatidylinositol 5-phosphate 4-kinase type-2 alpha IHC Antibodies

Anti-PIP4K2A antibodies A04244 and A04244-1 have IHC images from paraffin sections and IF images from cells (catalog image captions). Listed reactivity spans human, mouse and rat (catalog application and reactivity lists).

Real IHC data Immunohistochemistry of paraffin-embedded rat kidney using PIP4K2A antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Anti-PIP4K2A Antibody

Cat # A04244

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Real IHC data Immunohistochemical analysis of paraffin-embedded H. brain section using PIP4K2A Antibody (C-term). A04244-1 was diluted at 1:25 dilution. A undiluted biotinylated goat polyvalent antibody was used as the secondary, followed by DAB staining.

Anti-PIP4K2A Antibody (C-term)

Cat # A04244-1

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A04244 has IHC images from paraffin-embedded rat kidney and mouse lung and an IF image from A549 cells (A04244 image captions). A04244-1 has an IHC image from a paraffin-embedded human brain section and an IF image from HeLa cells (A04244-1 image captions).

Which to pick: For paraffin-section IHC, choose A04244 for rat kidney or mouse lung examples, or A04244-1 for a human brain example; both IHC captions leave the fixative unreported (A04244 and A04244-1 IHC image captions). For IF/ICC, A04244 lists both IF and ICC and shows IF in A549 cells, while A04244-1 lists IF and shows IF in HeLa cells (catalog application lists; A04244 and A04244-1 IF image captions). For cross-species work, A04244 lists human, mouse and rat reactivity, while A04244-1 lists human only (catalog reactivity lists).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P48426 (PI42A_HUMAN, Phosphatidylinositol 5-phosphate 4-kinase type-2 alpha).
Human Protein Atlas. PIP4K2A tissue IHC expression (reliability: Supported).
Human Protein Atlas. PIP4K2A subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the plasma membrane and cytosol..
Human Protein Atlas. PIP4K2A antibody validation summary (2 antibodies).
Exploring new mechanisms in cancer molecular pathways and pathogenic cell transformation: PIP4K2A as a prognostic marker and therapeutic target in cutaneous malignant melanoma. Discover oncology 2024 — PMC11585527.
PIP4K2A as a negative regulator of PI3K in PTEN-deficient glioblastoma. The Journal of experimental medicine 2019 — PMC6504209.
Loss of PI5P4Kα Slows the Progression of a Pten Mutant Basal Cell Model of Prostate Cancer. Molecular cancer research : MCR 2025 — PMC7616865.
PIP4K2A/2B inhibitor suppresses tumor growth in a xenograft model of NSCLC. iScience 2026 — PMC13206749.
PubMed PMID:7852364 — UniProt-cited evidence.
PubMed PMID:7639683 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.

Immunohistochemistry protocol sheet for PIP4K2A (Phosphatidylinositol 5-phosphate 4-kinase type-2 alpha). Expected localisation General nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A04244 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A04244); 2. Fixation: Image fixative and duration unreported (datasheet A04244); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 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-PIP4K2A, 1:50-1:200 (datasheet A04244); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep 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 A04244); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage for each (UniProt); Regulation: Brain-enhanced RNA expression (HPA tissue RNA). Evidence: UniProt P48426, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A04244, PMC11585527, PMC6504209. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PIP4K2A/immunohistochemistry-ihc-guide.