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PLA2G1B / Phospholipase A2 · IHC design guide
Design Immunohistochemistry for PLA2G1B
Plan PLA2G1B paraffin IHC using exocrine pancreas as a positive tissue and cytoplasmic glandular staining as the expected pattern (HPA tissue IHC). Compare consistently fixed sections and account for secretion when interpreting staining beyond the producing cells (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PLA2G1B 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 exocrine pancreas (HPA tissue IHC) | |
| Staining pattern | Exocrine glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Pancreas | |
| 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 | Secretion can separate protein from its source cells (HPA tissue IHC) | |
| Regulation | Pancreas-enriched expression (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms; residues 1–22 precede the mature chain (UniProt) |
Section 1
Recommended PLA2G1B IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published mouse lung IHC workflow (PMC12571731).
| Sample | Paraffin-embedded mouse pancreas tissue; fixative not specified (datasheet A06738-1) |
| Fixation | Image fixative and duration unreported (datasheet A06738-1); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-PLA2G1B, 1:100-1:200 (datasheet A06738-1) |
| 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 | PLA2G1B-positive staining in exocrine glandular cells of pancreas (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in exocrine pancreas. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the published EDTA sequence is an alternative (PMC12571731).
Section 2
What Is the Expected PLA2G1B Staining Pattern?
PLA2G1B is secreted from pancreatic acinar cells in an inactive form and has no transmembrane segment (UniProt P04054). In tissue IHC, expect selective cytoplasmic staining in pancreatic exocrine glandular cells (HPA: High in exocrine glandular cells). HPA rates this tissue pattern Enhanced, while cautioning that secretion can separate the locations of RNA and protein (HPA: Enhanced reliability).
| Strong cytoplasmic staining in pancreatic exocrine glandular cells, with little staining elsewhere on the section. | This matches the reported selective pancreatic pattern and High staining level (HPA: tissue IHC). Interpret the stained cells and compartment together: the HPA finding supports a cytoplasmic exocrine pattern, while UniProt identifies the protein as secreted (UniProt P04054). |
| Prominent nuclear staining, especially when cytoplasmic exocrine staining is weak or absent. | A dominant nuclear pattern conflicts with HPA's selective cytoplasmic observation and UniProt's secreted annotation (HPA: tissue IHC; UniProt P04054). Treat it as suspect staining; compare a no-primary control and reassess detection and image interpretation before assigning it to PLA2G1B (general IHC practice). |
| Strong staining in unrelated cell types, or in adipocytes or adrenal glandular cells. | HPA reports PLA2G1B as Not detected in those two listed cell types, so staining there warrants a specificity check (HPA: tissue IHC). Cross-reactivity or endogenous chromogenic activity are possible explanations, not diagnoses from appearance alone; compare controls and the expected pancreatic pattern (general IHC practice). |
| Diffuse color across multiple compartments, including areas without a recognizable exocrine cell pattern. | Uniform haze does not reproduce HPA's selective cytoplasmic exocrine result (HPA: tissue IHC). Check no-primary controls, endogenous detection activity, blocking and chromogen development before scoring cells; widespread color can obscure a genuine localized signal (general IHC practice). |
| No convincing cytoplasmic signal in a pancreatic exocrine region. | That conflicts with the High pancreatic exocrine staining reported by HPA, but one negative section does not establish biological absence (HPA: tissue IHC). Confirm that exocrine tissue is present, then assess the assay with a suitable positive control and review antibody and detection conditions (general IHC practice). |
💡Expected PLA2G1B appearanceCall a result positive when pancreatic exocrine glandular cells show High, selective cytoplasmic staining; dominant nuclear color or broad staining of unrelated cells is suspect (HPA: tissue IHC).
| Secreted protein and processing | UniProt describes secretion from pancreatic acinar cells, a signal peptide at residues 1–15, a propeptide at 16–22, and a mature chain at 23–148 (UniProt P04054). An extracellular signal should therefore be assessed in context; secretion alone does not make diffuse slide background specific staining. |
| Tissue context | HPA calls pancreatic RNA tissue enriched and pancreatic exocrine IHC High, while its listed samples include several Not detected cell types (HPA: tissue IHC). UniProt also reports expression in lung, liver and kidney (UniProt P04054). Do not infer that every nonpancreatic tissue must be IHC negative. |
| Antibody evidence | Two rabbit polyclonal antibodies, HPA047822 and HPA060803, have Enhanced IHC status (HPA: antibodies). That supports the reported tissue pattern; it does not establish that every new antibody, staining run or unexpected compartment is specific (general IHC practice). |
| Can the IHC pattern be applied to ICC/IF? | Use the pancreatic IHC pattern as context, not as an established cellular IF image: HPA lists no main subcellular location or ICC/IF image cell lines for PLA2G1B (HPA: subcellular). UniProt calls it secreted (UniProt P04054). Interpret any ICC/IF signal with its own controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| Pancreatic exocrine cells are unstained. | The result conflicts with HPA's High exocrine signal; the reason cannot be assigned from the section alone (HPA: tissue IHC). | Verify exocrine tissue, positive control performance, antibody application and detection; review retrieval conditions without assuming PLA2G1B-specific fixation sensitivity (general IHC practice). |
| The whole section has weak, even color. | Diffuse color lacks the selective cytoplasmic distribution reported by HPA; detection background is possible (HPA: tissue IHC; general IHC practice). | Examine a no-primary control; review blocking, washes, endogenous activity control and chromogen development before scoring (general IHC practice). |
| Nuclei dominate the signal. | A dominant nuclear pattern disagrees with the HPA cytoplasmic observation and UniProt secreted annotation (HPA: tissue IHC; UniProt P04054). | Check counterstain separation and no-primary controls, then confirm whether cytoplasmic exocrine staining is independently visible (general IHC practice). |
| Adipocytes or adrenal glandular cells stain strongly. | HPA lists both cell types as Not detected; cross-reactivity or endogenous detection activity should be investigated (HPA: tissue IHC; general IHC practice). | Compare negative and no-primary controls with pancreatic positive tissue; do not score the unexpected color as PLA2G1B without corroboration (general IHC practice). |
| Extracellular color is present near otherwise plausible pancreatic staining. | Secretion makes extracellular protein plausible, but its exact histologic distribution is not established by the supplied HPA pattern (UniProt P04054; HPA: tissue IHC). | Score the documented cytoplasmic exocrine pattern separately; use controls to distinguish localized signal from diffuse chromogenic background (HPA: tissue IHC; general IHC practice). |
| An ICC/IF image appears unlike the pancreatic IHC section. | HPA provides no PLA2G1B ICC/IF image cell lines or main subcellular location for a direct comparison (HPA: subcellular). | Interpret ICC/IF with application-specific positive and negative controls; avoid transferring an IHC staining claim to a cellular IF result (general IHC/IF practice). |
Experiment evidence
IHC & IF images showing PLA2G1B staining
A06738-1 · IHCImmunohistochemistry of paraffin-embedded mouse pancreas using PLA2G1B antibody at dilution of 1:100 .Perform microwave antigen retrieval wi
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000170890-PLA2G1B/tissue/cerebral+cortex#imid_16723331
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000170890-PLA2G1B/tissue/colon#imid_16723761
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000170890-PLA2G1B/tissue/liver#imid_16723770
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000170890-PLA2G1B/tissue/pancreas#imid_16723721
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PLA2G1B IHC & IF
🧪Run pancreas first: exocrine glandular cells should stain strongly (HPA: High in pancreatic exocrine glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); cells outside the exocrine compartment on the pancreas slide should provide a low-background comparison, without assuming every such cell is target-negative.
Positive control tissue: Pancreas (Exocrine glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PLA2G1B; derive a cell-line control from the positive tissue's cell type (Exocrine glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an irrelevant immunoglobulin matched to the primary antibody’s host species, isotype and clonality; and PLA2G1B knockout tissue as a biological negative. Quench endogenous peroxidase for chromogenic IHC, and assess pancreatic autofluorescence separately if using IF.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (A06738-1 caption). The caption uses microwave antigen retrieval in 10 mM PBS at pH 7.2 and a 1:100 antibody dilution; compare a matched section without retrieval to assess dependence (A06738-1 caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; extracellular staining may complicate cell attribution because PLA2G1B is secreted from pancreatic acinar cells (UniProt P04054: Secreted).
HPA tissue IHC evidence for PLA2G1B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PLA2G1B IHC Tips
Troubleshoot PLA2G1B staining in chromogenic IHC using the documented pancreas pattern, secreted topology, and antibody-specific processing evidence.
How should I adjust retrieval when PLA2G1B staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page antigen-retrieval rule). If staining is weak, compare a separate section retrieved by microwave in 10 mM PBS, pH 7.2, the condition reported for catalog antibody A06738-1 in paraffin-embedded mouse pancreas (A06738-1 tissue-IHC caption). Run the same positive-control tissue through both conditions and compare signal in exocrine glandular cells, where cytoplasmic expression is expected (HPA tissue IHC). Keep antibody concentration and chromogen development constant across the comparison so retrieval is the main changed variable (standard IHC practice).
Could fixation explain weak or patchy PLA2G1B IHC staining?
Target-specific sensitivity to fixation is unknown because the supplied A06738-1 pancreas caption identifies paraffin sections but does not state the fixative (A06738-1 tissue-IHC caption). Record the actual fixative, fixation duration, and processing history for each specimen before attributing a weak result to antigen loss (standard IHC practice). Compare sections processed together, using exocrine pancreas as a positive reference because that is the documented high-staining compartment (HPA tissue IHC). If fixation histories differ, repeat the comparison with matched processing and the same retrieval and detection settings; a staining difference alone cannot establish a PLA2G1B-specific fixation effect (standard IHC practice).
Where should PLA2G1B signal appear, and how should extracellular staining be judged?
Expect a selective cytoplasmic pattern in pancreatic exocrine glandular cells in tissue IHC (HPA tissue IHC). PLA2G1B is secreted from pancreatic acinar cells in an inactive form and has no transmembrane segment, so extracellular signal can be biologically plausible but needs careful assignment to tissue structures (UniProt P04054 subcellular annotation and topology). Compare cell-associated and extracellular staining on the same section, including the no-primary control, before calling a diffuse deposit specific (standard IHC practice). Do not interpret a crisp plasma-membrane outline as proof that PLA2G1B is an integral membrane protein; receptor interaction and extracellular clearance are separately reported properties (UniProt P04054 topology and subunit annotation).
How do precursor processing and epitope placement affect interpretation of PLA2G1B staining?
Check the antibody's disclosed immunogen or epitope against the 1–15 signal peptide, 16–22 propeptide, and 23–148 mature chain before comparing staining across specimens (UniProt P04054 processing annotation). Processing could remove an epitope in either N-terminal segment, so staining intensity may reflect epitope availability as well as protein abundance if the antibody targets that segment (UniProt P04054 processing annotation; standard IHC interpretation). No isoforms, glycosylation sites, or modified residues are annotated in the supplied record; those mechanisms should not be assumed to explain a pattern (UniProt P04054 record). If the epitope is undisclosed, report that limitation and validate any processing-dependent interpretation with an independently characterized reagent (standard IHC practice).
How should I adapt this PLA2G1B IHC result for multiplex IF?
Treat IF as a separate antibody validation because the supplied A06738-1 evidence describes paraffin-section IHC, and the HPA subcellular record lists no ICC/IF images (A06738-1 tissue-IHC caption; HPA subcellular). Multiplex PLA2G1B with a validated marker of pancreatic exocrine glandular cells to test whether cellular signal matches the expected tissue compartment (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores after inspecting tissue autofluorescence, and include single-stain and no-primary controls to distinguish bleed-through from specific signal (standard IF practice). For an intracellular epitope, compare gentle permeabilisation with an unpermeabilised control; epitope position must be checked because PLA2G1B is secreted and lacks a transmembrane segment (UniProt P04054 topology; standard IF practice).
What controls help distinguish PLA2G1B signal from chromogenic IHC background?
Inspect a no-primary control for staining from the detection system, and apply the routine endogenous-peroxidase block when using peroxidase and DAB (standard chromogenic IHC practice). Compare background in low-expected-signal reference areas with exocrine pancreatic cells, where high PLA2G1B staining is documented (HPA tissue IHC). If staining is widespread, titrate the antibody around the caption's 1:100 dilution while holding retrieval and DAB development constant (A06738-1 tissue-IHC caption; standard IHC practice). Evaluate section edges, folds, damaged areas, and precipitate separately from intact cells before changing the biological interpretation of a diffuse signal (standard IHC practice).
How should PLA2G1B IHC staining be scored across pancreatic sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the exocrine glandular cell compartment before scoring because it is the documented site of high cytoplasmic staining (HPA tissue IHC). Record the percentage of positive cells and intensity categories, then calculate an H-score consistently across specimens if intensity is informative (standard IHC quantification practice). Normalise counts to the number of evaluable exocrine cells, or normalise stained area to evaluable exocrine tissue area, rather than total section area when tissue composition varies (standard IHC quantification practice). Keep retrieval, antibody dilution, imaging, and chromogen development matched, and exclude folds, edges, and necrotic areas under a prespecified rule (standard IHC quantification practice).
When is an unexpected PLA2G1B staining pattern more likely to be artefact?
A convincing result should include cytoplasmic staining in pancreatic exocrine glandular cells, the documented high-expression compartment (HPA tissue IHC). Signal confined to an unrelated cell type, a section edge, or necrotic material deserves review against morphology and controls before it is called PLA2G1B (HPA tissue IHC; standard IHC interpretation). A membrane-only pattern is inconsistent with treating PLA2G1B as a transmembrane protein, although extracellular protein may associate with receptors or be cleared by endocytosis (UniProt P04054 topology and subunit annotation). Check no-primary and peroxidase-block controls for endogenous enzyme or detection-related colour, then repeat questionable staining in an intact, well-processed section (standard chromogenic IHC practice).
Boster reagents
Best PLA2G1B / Phospholipase A2 IHC Antibodies
PLA2G1B antibodies list human and mouse reactivity (catalog: both SKUs). A06738-1 has a mouse pancreas paraffin IHC figure; A06738-2 lists IF, with no IF figure provided (catalog image captions; applications).
Real IHC data
Anti-Phospholipase A2 PLA2G1B Antibody
Cat # A06738-1
A06738-1 lists human and mouse IHC reactivity and shows staining of mouse pancreas paraffin sections at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2 (catalog applications/reactivity; A06738-1 image caption). A06738-2 lists human and mouse IHC and IF reactivity, with no IHC or IF image caption provided (catalog applications/reactivity; image captions).
Which to pick: For tissue IHC, choose A06738-1 when a pictured mouse pancreas paraffin example is useful; its caption does not report the fixative (A06738-1 image caption). For IF, choose A06738-2 because IF is listed, while ICC is unlisted and no IF figure is provided (catalog applications; image captions). Both SKUs list human and mouse reactivity, but the pictured tissue result is mouse only (catalog reactivity; A06738-1 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P04054 (PA21B_HUMAN, Phospholipase A2).
- Human Protein Atlas. PLA2G1B tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PLA2G1B subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. PLA2G1B antibody validation summary (2 antibodies).
- Integrating bioinformatics and molecular experiments to reveal the critical role of the cellular energy metabolism-related marker PLA2G1B in COPD epithelial cells. Frontiers in immunology 2025 — PMC12571731.
- Microbial Protein Binding to gC1qR Drives PLA2G1B-Induced CD4 T-Cell Anergy. Frontiers in immunology 2022 — PMC8981723.
- PubMed PMID:3028739 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PLA2G1B (Phospholipase A2). Expected localisation Cytoplasmic staining in exocrine pancreas (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06738-1 at 1:100-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Pancreas (Exocrine glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse pancreas tissue; fixative not specified (datasheet A06738-1); 2. Fixation: Image fixative and duration unreported (datasheet A06738-1); 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 citrate buffer, pH 6.0 , 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-PLA2G1B, 1:100-1:200 (datasheet A06738-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secretion can separate protein from its source cells (HPA tissue IHC); Isoform / epitope: No annotated isoforms; residues 1–22 precede the mature chain (UniProt); Regulation: Pancreas-enriched expression (HPA tissue IHC). Evidence: UniProt P04054, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A06738-1, PMC12571731. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PLA2G1B/immunohistochemistry-ihc-guide.