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PLA2G4A / Cytosolic phospholipase A2 · IHC design guide
Design Immunohistochemistry for PLA2G4A
Plan PLA2G4A IHC-P around cytoplasmic tissue staining, with high staining in parathyroid and seminal-vesicle glandular cells (HPA tissue IHC). Start with the catalog antibody’s 1:10–1:50 IHC-P dilution range (datasheet), and consider calcium-dependent membrane translocation when interpreting compartment changes (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLA2G4A 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 tissue staining (HPA tissue IHC); Golgi and nuclear-envelope association possible (UniProt) | |
| Staining pattern | Cytoplasmic; high in parathyroid and seminal-vesicle glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Parathyroid gland+4 more · see all | |
| Negative control | Ovary+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Low antibody–RNA concordance limits tissue predictions (HPA tissue IHC) | |
| Regulation | Intensity regulation not specified (UniProt) | |
| Isoform / epitope | No listed isoforms; one 1–749 chain with no transmembrane segment (UniProt) |
Section 1
Recommended PLA2G4A IHC & IF Protocols
The catalog antibody protocol is paired with a published chromogenic IHC protocol for mouse testis (PMC4372093).
| Sample | FFPE human lung carcinoma tissue (datasheet A00854-1) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A00854-1); 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-PLA2G4A, 1:10-1:50 (datasheet A00854-1) |
| 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 | PLA2G4A-positive staining in glandular cells of parathyroid gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in parathyroid gland and seminal vesicle. No signal in the no-primary control. |
💡Decision noteTry Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min first (page antigen-retrieval setting); the published protocol does not report retrieval (PMC4372093).
Section 2
What Is the Expected PLA2G4A Staining Pattern?
PLA2G4A should stain mainly in the cytoplasm, with possible staining at intracellular membranes and the nuclear envelope; it has no transmembrane segment (UniProt P47712). In tissue IHC, glandular cells of parathyroid gland and seminal vesicle show high staining (HPA tissue IHC). HPA rates its tissue staining Approved but reports low consistency with RNA expression, so interpret unexpected patterns cautiously (HPA tissue IHC).
| Cytoplasmic staining in parathyroid or seminal vesicle glandular cells. | This matches the high staining reported for those cells (HPA tissue IHC) and the principal cytoplasmic localization of PLA2G4A (UniProt P47712). Assess the named cells rather than assigning the same intensity to every cell in the section; HPA reports a cell-specific tissue pattern (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear envelope component is possible, but predominantly intranuclear staining is outside the reported main cytoplasmic pattern (UniProt P47712; HPA subcellular ICC-IF). Check morphology and counterstain before assigning a compartment (standard IHC practice). An unusual compartment alone does not establish target specificity, especially given HPA's low staining-to-RNA consistency (HPA tissue IHC). |
| Strong signal in ovarian stroma, salivary glandular cells, or vaginal squamous epithelium. | HPA reports PLA2G4A as not detected in those respective cells (HPA tissue IHC). Consider cross-reactivity or endogenous chromogen-generating activity, and inspect a detection-only control (standard IHC practice). Treat these tissues as comparison areas, not absolute biological negatives: HPA notes low consistency between its antibody staining and RNA expression data (HPA tissue IHC). |
| Uniform color across tissue, including empty spaces or unrelated structures. | That distribution is difficult to reconcile with the reported cell-associated cytoplasmic pattern (HPA tissue IHC; UniProt P47712). Diffuse deposition can reflect incomplete blocking, residual detection activity, or overly strong detection chemistry (standard IHC practice). Compare with a detection-only control before scoring cells as positive (standard IHC practice). |
| No signal in parathyroid or seminal vesicle glandular cells. | These are high-staining reference cells in the HPA tissue survey (HPA tissue IHC). First verify that the expected cells are present and that counterstain and detection worked (standard IHC practice). Then review the antibody's documented IHC conditions and the run controls; absence of color in one section does not establish absence of PLA2G4A (standard IHC practice). |
💡Expected PLA2G4A appearanceCall a convincing positive when glandular cells show clear cytoplasmic staining, potentially with intracellular membrane accentuation, and strongest expected staining in parathyroid gland or seminal vesicle (UniProt P47712; HPA tissue IHC); indiscriminate background or dominant intranuclear color is suspect (standard IHC practice; HPA subcellular ICC-IF).
| Compartment and cell state | PLA2G4A is cytoplasmic and can translocate to intracellular membranes in a calcium-dependent way (UniProt P47712). Membrane-associated accentuation can therefore coexist with cytoplasmic signal; a single fixed section does not reveal when translocation occurred (UniProt P47712; standard IHC interpretation). |
| Tissue and cell selection | HPA reports high glandular staining in parathyroid gland and seminal vesicle, medium staining in bronchial respiratory epithelium, and no detected staining in ovarian stroma (HPA tissue IHC). Compare matched cell types when judging intensity; whole-tissue color can obscure that distinction (standard IHC practice). |
| Evidence strength | The HPA tissue profile is Approved but has low consistency between antibody staining and RNA expression (HPA tissue IHC). Its listed antibodies have Approved IHC status, which supports use of the reported pattern while leaving an unexpected result in need of controls (HPA antibodies; standard IHC practice). |
| Protein architecture | UniProt lists no signal peptide, propeptide, transmembrane segment, glycosylation site, or isoform for the supplied record (UniProt P47712). These annotations give no basis here for expecting an extracellular or shed-protein staining pattern (UniProt P47712). The antibody epitope is unspecified, so epitope-specific retrieval predictions are unsupported. |
| IF/ICC Q&A: where should signal appear? | Mainly in the cytosol, with additional vesicular signal (HPA subcellular ICC-IF). UniProt also lists Golgi membrane and nuclear envelope localization (UniProt P47712). This answers the localization question only; the supplied evidence gives no IF/ICC protocol conditions. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-reference glandular cells are blank. | The expected cells may be absent from the section, or an IHC run step may have failed (HPA tissue IHC; standard IHC practice). | Locate glandular cells, check a run control, and review the antibody's documented IHC conditions before interpreting the blank section (standard IHC practice). |
| Color is strongest in nuclei. | The compartment differs from the mainly cytoplasmic pattern; nuclear envelope localization should not be read as diffuse intranuclear staining (UniProt P47712; HPA subcellular ICC-IF). | Recheck focus and counterstain, then compare the signal with a detection-only control (standard IHC practice). |
| Reported negative cells stain strongly. | Cross-reactivity or endogenous detection activity is possible; HPA reports no detection in ovarian stroma, salivary glandular cells, and vaginal squamous epithelium (HPA tissue IHC; standard IHC practice). | Inspect cell identity and a detection-only control; do not assign target-specific positivity from color alone (standard IHC practice). |
| The entire section has diffuse chromogen. | Nonspecific binding, incomplete blocking, or excessive detection signal can obscure cell boundaries (standard IHC practice). | Compare the detection-only control and assess blocking, washing, and detection conditions using the documented workflow (standard IHC practice). |
| Cytoplasm stains, but intensity differs among tissues. | HPA records high, medium, low, and undetected cell-specific levels across tissues, with low staining-to-RNA consistency (HPA tissue IHC). | Score the named cell population and compartment against appropriate controls; avoid treating one tissue's intensity as a universal threshold (HPA tissue IHC; standard IHC practice). |
| Puncta or membrane-associated color accompany cytoplasmic signal. | Calcium-dependent movement to intracellular membranes is reported for PLA2G4A, while ICC-IF also supports vesicular localization (UniProt P47712; HPA subcellular ICC-IF). | Record the distribution alongside cytoplasmic staining and morphology; use controls before deciding whether the puncta are specific (standard IHC practice). |
Experiment evidence
IHC & IF images showing PLA2G4A staining
A00854-1 · IHCFormalin-fixed and paraffin-embedded human lung carcinoma with PLA2G4A Antibody (Center), which was peroxidase-conjugated to the secondary a
A00854-1 · IFConfocal immunofluorescent analysis of PLA2G4A Antibody (Center) with NCI-H460 cell followed by Alexa Fluor® 488-conjugated goat anti-rabbit
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000116711-PLA2G4A/tissue/cerebral+cortex#imid_3126360
HPA · Parathyroid glandHuman Protein Atlas IHC, Parathyroid gland. Source: https://www.proteinatlas.org/ENSG00000116711-PLA2G4A/tissue/parathyroid+gland#imid_3126251
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000116711-PLA2G4A/tissue/colon#imid_3126286
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000116711-PLA2G4A/tissue/liver#imid_3126287
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA050062; green = PLA2G4A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA050062; green = PLA2G4A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLA2G4A IHC & IF
🧪Run parathyroid gland first: its glandular cells should stain (HPA: High in parathyroid gland glandular cells). Use salivary gland glandular cells as the negative tissue (HPA: Not detected); on the parathyroid slide, assess nonglandular cells for little or no specific staining as an internal negative (standard IHC practice).
Positive control tissue: Parathyroid gland (Glandular cells, HPA High)
Negative control tissue: Ovary (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PLA2G4A in A-549, HeLa, NIH 3T3, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and a PLA2G4A knockout sample or validated peptide-block control if available (standard IHC practice). Check endogenous peroxidase background in the parathyroid section before interpreting DAB staining (standard chromogenic IHC practice).
⚠️Feasibility: The selected A00854-1 image caption reports a formalin-fixed, paraffin-embedded lung carcinoma specimen stained with DAB (selected-SKU IHC caption); it reports no target-specific fixation window, and retrieval dependency is unreported in the supplied evidence. Whether frozen sections or IF are easier is unreported in the supplied evidence. Assess background in the parathyroid glandular cells used as the positive readout before scoring PLA2G4A staining (HPA: High in parathyroid gland glandular cells; standard IHC practice).
HPA tissue IHC evidence for PLA2G4A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PLA2G4A IHC Tips
Troubleshoot PLA2G4A staining in paraffin sections by checking retrieval, cell type, and compartment before comparing chromogenic IHC results (UniProt P47712; HPA tissue IHC).
How should I adjust retrieval when PLA2G4A staining is weak?
Start with Tris-EDTA at pH 9.0 for heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). If staining remains weak, compare a small section series with citrate at pH 6.0 as a fallback, keeping antibody concentration and detection conditions constant (standard IHC practice). Record section loss, tissue morphology, and staining in the expected cytoplasmic compartment, since PLA2G4A is mainly cytoplasmic and can associate with intracellular membranes (UniProt P47712 subcellular location). Use the same retrieval run for comparison sections and controls so differences in heating do not masquerade as biological differences (standard IHC practice).
Could fixation explain absent or uneven PLA2G4A staining?
Target-specific fixation sensitivity for PLA2G4A is unknown from the supplied evidence; assess fixation as a general section-quality variable (supplied fixation rule; standard IHC practice). Compare sections with documented processing histories, then check morphology, section adhesion, and staining across the full tissue area before changing antibody conditions (standard IHC practice). For a routine formalin-fixed, paraffin-embedded workflow, keep deparaffinisation, retrieval, and detection consistent across the comparison (standard IHC practice). Do not infer a PLA2G4A-specific fixation effect from its cytoplasmic and membrane-associated locations or phosphorylation sites, which describe the protein rather than its response to fixation (UniProt P47712 location and modified residues).
Is perinuclear or punctate staining compatible with PLA2G4A localisation?
Yes: PLA2G4A is cytoplasmic and can translocate to intracellular membranes in a calcium-dependent manner, including the Golgi apparatus membrane and nuclear envelope (UniProt P47712 subcellular location). Mainly cytosolic staining with additional vesicular signal is also reported in cell imaging (HPA subcellular: supported cytosol and vesicles). Evaluate perinuclear signal against cell boundaries and a nuclear counterstain; a nuclear rim differs from diffuse staining throughout the nucleus (UniProt P47712 subcellular location; standard IHC practice). Because the record lists no transmembrane segment, uniform cell-surface outlining warrants review of section background and detection controls before assigning it to PLA2G4A (UniProt P47712 topology; standard IHC practice).
How can epitope choice affect PLA2G4A staining in paraffin sections?
Map the antibody’s stated immunogen to PLA2G4A before interpreting regional staining: the record identifies a C2 domain at residues 6–122 and a PLA2c domain at 140–740 (UniProt P47712 domains). The supplied record lists 0 isoforms, so do not explain discordant staining by an isoform without independent evidence (UniProt P47712 isoforms). Several modified residues are reported, including phosphosites at 268 and 434–437, but the payload does not locate this antibody’s epitope (UniProt P47712 modified residues; supplied antibody evidence). If retrieval changes staining, compare morphology and controls alongside signal; neither epitope masking nor phosphorylation dependence is established for this antibody (standard IHC practice; supplied antibody evidence).
How should I investigate PLA2G4A localisation by IF alongside this IHC result?
For a separate IF experiment, pair PLA2G4A with a validated macrophage marker when macrophages are the cells under study; macrophages are among the reported expressing cells (UniProt P47712 tissue specificity). Choose well-separated fluorophores and inspect unstained tissue for autofluorescence before assigning weak puncta to PLA2G4A (standard IF practice). Optimise mild permeabilisation for an intracellular epitope, since PLA2G4A has no transmembrane segment and occupies cytosolic and intracellular membrane-associated locations (UniProt P47712 topology and subcellular location; standard IF practice). Compare the resulting cytosolic and vesicular pattern with IF localisation evidence, while interpreting the chromogenic paraffin-section result on its own controls (HPA subcellular: supported cytosol and vesicles; standard IHC/IF practice).
What causes diffuse brown background in PLA2G4A IHC?
First inspect a no-primary control and tissue edges to separate detection background from the general cytoplasmic PLA2G4A pattern reported in tissue (standard IHC practice; HPA tissue IHC). In a peroxidase/DAB workflow, verify the peroxide block, adequate washes, and controlled DAB development, since residual endogenous peroxidase or overdevelopment can create brown signal (standard IHC practice). Check blocking and titrate the catalog antibody only within conditions supported by its instructions; no dilution is provided in this payload (supplied antibody evidence; standard IHC practice). Judge improvement by cleaner negative compartments and preserved staining in expected cells, rather than by reduced overall colour alone (UniProt P47712 tissue specificity and subcellular location; standard IHC practice).
How should I score PLA2G4A staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because PLA2G4A can be cytosolic or associated with intracellular membranes (UniProt P47712 subcellular location). For a defined cell population, report the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories and their cell percentages (standard IHC practice). If measuring stained cells per mm², normalise to viable tissue area and report the number of analysed cells or fields (standard IHC practice). Keep retrieval, imaging, threshold, and counterstain settings consistent, and stratify cell types because expression differs across tissues and cells (standard IHC practice; HPA tissue IHC).
How can I distinguish genuine PLA2G4A signal from artefact?
Look for staining in plausible cells and compartments: PLA2G4A is reported in macrophages and other cells, with cytoplasmic and intracellular membrane-associated localisation (UniProt P47712 tissue specificity and subcellular location). A no-primary control helps identify endogenous peroxidase or detection background, while edge-only or necrotic-area staining should be reviewed against tissue morphology (standard IHC practice). Interpret a strong signal in an unexpected compartment cautiously and check the nuclear counterstain and adjacent intact cells (UniProt P47712 subcellular location; standard IHC practice). The selected antibody has a formalin-fixed, paraffin-embedded human lung carcinoma DAB image, but that caption does not establish clinical relevance or prove every stained cell is specific (A00854-1 tissue-IHC caption).
Boster reagents
Best PLA2G4A / Cytosolic phospholipase A2 IHC Antibodies
Human-reactive A00854-1 has IHC data from formalin-fixed, paraffin-embedded human lung carcinoma and IF data from NCI-H460 cells (catalog: reactivity; IHC and IF image captions).
Real IHC data
Anti-PLA2G4A Antibody (Center)
Cat # A00854-1
A00854-1 is listed for human IHC-P, with a DAB-stained image of formalin-fixed, paraffin-embedded human lung carcinoma (catalog: applications, reactivity, IHC image caption). The same SKU is listed for IF and has a confocal IF image of NCI-H460 cells (catalog: applications, IF image caption).
Which to pick: Choose A00854-1 for human tissue IHC-P: its own image documents formalin-fixed, paraffin-embedded lung carcinoma, and its listed IHC-P dilution is 1:10–1:50 (catalog: IHC image caption, reactivity, IHC-P dilution). For IF/ICC planning, A00854-1 is a rabbit polyclonal antibody with listed IF use and an NCI-H460 IF image; ICC is not separately listed (catalog: host, dilution_raw, applications, IF image caption). No cross-species option is supported because A00854-1 lists human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P47712 (PA24A_HUMAN, Cytosolic phospholipase A2).
- Human Protein Atlas. PLA2G4A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PLA2G4A subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to vesicles..
- Human Protein Atlas. PLA2G4A antibody validation summary (3 antibodies).
- Multi-omics analysis of the effects of pla2g4a on the prognosis of various cancers and its experimental validation in breast cancer cell lines. Discover oncology 2025 — PMC12234970.
- Group IVA phospholipase A2 regulates testosterone biosynthesis by murine Leydig cells and is required for timely sexual maturation. The Biochemical journal 2011 — PMC4372093.
- scRNA-seq reveals that VEGF signaling mediates the response to neoadjuvant anlotinib combined with PD-1 blockade therapy in non-small cell lung cancer. Journal of translational medicine 2025 — PMC12032801.
- Effects of Clinacanthus nutans Extracts on Cell Proliferation and Apoptosis in Triple-Negative Breast Cancer: Mechanistic Insights. Breast cancer (Dove Medical Press) 2025 — PMC12448096.
- PubMed PMID:1904318 — UniProt-cited evidence.
- PubMed PMID:1869522 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLA2G4A (Cytosolic phospholipase A2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); Golgi and nuclear-envelope association possible (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A00854-1 at 1:10-1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Parathyroid gland (Glandular cells); negative Ovary. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: FFPE human lung carcinoma tissue (datasheet A00854-1); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A00854-1); 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-PLA2G4A, 1:10-1:50 (datasheet A00854-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Low antibody–RNA concordance limits tissue predictions (HPA tissue IHC); Isoform / epitope: No listed isoforms; one 1–749 chain with no transmembrane segment (UniProt); Regulation: Intensity regulation not specified (UniProt). Evidence: UniProt P47712, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to vesicles., Datasheet A00854-1, PMC4372093. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLA2G4A/immunohistochemistry-ihc-guide.