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PON2 / Serum paraoxonase/arylesterase 2 · IHC design guide
Design Immunohistochemistry for PON2
Plan PON2 paraffin-section IHC around its generally cytoplasmic tissue staining (HPA tissue IHC) and membrane annotation (UniProt). Use high-staining glandular or glial cells and tissues with undetected staining as reference controls (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PON2 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 cytoplasmic tissue staining (HPA tissue IHC); membrane annotated (UniProt) | |
| Staining pattern | High staining in glandular and glial cells; generally cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet M02027-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Esophagus+2 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 | A secreted variant may shift protein from RNA-defined sites (HPA tissue IHC) | |
| Regulation | Widely expressed; tissue levels vary (UniProt) | |
| Isoform / epitope | 3 isoforms; verify which forms the epitope covers (UniProt) |
Section 1
Recommended PON2 IHC & IF Protocols
The catalog antibody protocol is paired with 3 published PON2 chromogenic IHC protocols for lymphoma, glioblastoma and pancreatic tissue (PMC9559328; PMC5362434; PMC5567863).
| Sample | Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet M02027-1) |
| Fixation | Image fixative and duration unreported (datasheet M02027-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0, 20 minutes (datasheet M02027-1) |
| 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 monoclonal (clone 19P05) anti-PON2, 1:50 (datasheet M02027-1) |
| 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 | PON2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA at pH 8.0 for the catalog antibody (datasheet M02027-1). The lymphoma protocol used citrate at pH 6.0 (PMC9559328).
Section 2
What Is the Expected PON2 Staining Pattern?
PON2 should show predominantly cytoplasmic staining, with possible membrane-associated signal, in glandular cells and glia (HPA: general cytoplasmic expression; HPA: high glandular and glial staining; UniProt Q15165: membrane, no transmembrane segment). HPA rates tissue IHC reliability Enhanced, while noting only medium consistency with RNA data (HPA: tissue IHC reliability description). Judge the pattern by cell type and compartment together, not by overall tissue darkness.
| Clear cytoplasmic staining in adrenal gland or duodenal glandular cells, or in cerebral cortex glia. | This fits the reported high cell-specific signal (HPA: High in these cells). Compare stained cells with adjacent structures; a uniformly dark section does not establish the reported pattern (HPA: general cytoplasmic expression). |
| Predominantly nuclear or other isolated compartment staining in otherwise positive cells. | This is discordant with the reported cytoplasmic pattern (HPA: general cytoplasmic expression). Review morphology, counterstain and detection controls before calling it PON2; UniProt's membrane annotation does not establish a nuclear pattern (UniProt Q15165: membrane). |
| Strong signal confined to esophageal or vaginal squamous epithelial cells, or salivary glandular cells. | Those cells were reported as not detected (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous detection activity, then check a positive tissue alongside them; an unexpected stain alone does not identify its cause. |
| Color spread across stroma, lumina and many unrelated cell types without distinct cellular outlines. | Treat this as diffuse background rather than a cell-specific positive result (HPA: general cytoplasmic expression). Check a control without primary antibody and inspect whether the background follows tissue structures or the detection chemistry (general IHC practice). |
| No convincing signal in adrenal glandular cells or cerebellar Bergmann glia. | Both have reported high staining (HPA: tissue IHC). First verify tissue identity, cell preservation and a working positive control; a negative slide may reflect assay performance and should not establish absence of PON2 by itself. |
💡Expected PON2 appearanceCall a result positive when cytoplasmic, sometimes membrane-associated color is strongest in the expected glandular cells or glia (HPA: High; UniProt Q15165: membrane); isolated nuclear color or diffuse cell-independent deposit is suspect (HPA: general cytoplasmic expression).
| Cell type within the section | HPA reports high staining in selected glandular cells and glia, but not detected staining in specified squamous or salivary cells (HPA: tissue IHC). Score the named cells rather than assigning one intensity to a whole organ. |
| Compartment assignment | HPA describes general cytoplasmic expression, while UniProt annotates membrane association without a transmembrane segment (HPA: tissue IHC; UniProt Q15165 topology). This supports a cytoplasmic or membrane-associated reading, not a requirement for a sharp surface rim. |
| Antibody validation | HPA029193 has Enhanced IHC validation and CAB025430 has Supported IHC validation (HPA: antibody validation). These grades help prioritize concordant patterns; they do not prove that every colored structure in a new specimen is PON2. |
| Isoforms and processing | Three isoforms and three glycosylation sites are annotated (UniProt Q15165: isoforms; glycosylation at 254, 269, 323). No supplied epitope map links these features to IHC staining, so do not predict a compartment or tissue difference from them. |
| IF/ICC evidence | HPA labels the subcellular summary Secreted but provides no main location or ICC-IF image-bearing cell lines (HPA: ICC-IF). Its tissue IHC description remains the direct basis for this paraffin-section call; an IF/ICC pattern needs its own validation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining glandular or glial cells look negative. | Tissue sampling, detection failure or assay conditions may be responsible; HPA reports these cells as High (HPA: tissue IHC). | Confirm cell identity and run a known-positive section with the same stain; review the IHC-validated antibody's documented IHC-P conditions (general IHC practice). |
| Every compartment is weak and poorly defined. | Diffuse background can obscure the expected cellular pattern (HPA: general cytoplasmic expression); its source cannot be assigned from the image alone. | Compare a no-primary control, then review blocking, washes and detection exposure for the entire run (general IHC practice). |
| Unexpected color appears in HPA not-detected cell types. | Cross-reactivity or endogenous detection activity is possible; HPA reports no detection in esophageal squamous and salivary glandular cells (HPA: tissue IHC). | Check a no-primary control and an expected positive tissue; interpret residual cell-specific staining cautiously (general IHC practice). |
| Only nuclei appear positive. | Nuclear-only staining conflicts with HPA's general cytoplasmic pattern (HPA: tissue IHC); the cause remains undetermined. | Recheck compartment boundaries against the counterstain and compare controls before scoring nuclei as PON2 (general IHC practice). |
| Two antibodies give different tissue patterns. | Their IHC validation levels differ: HPA029193 is Enhanced and CAB025430 is Supported (HPA: antibody validation). This does not by itself explain the disagreement. | Compare the same cell types on matched sections and retain the discordance in the interpretation (general IHC practice). |
| Can an IF/ICC image establish the expected IHC-P pattern? | HPA supplies no ICC-IF image-bearing cell lines or main subcellular location for PON2 (HPA: ICC-IF). | Use HPA tissue IHC to assess paraffin sections; evaluate IF/ICC separately with its own controls (HPA: tissue IHC; general IF practice). |
Experiment evidence
IHC & IF images showing PON2 staining
M02027-1 · IHCImmunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-PON2 antibody. The section was pre-treated using h
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105854-PON2/tissue/cerebral+cortex#imid_6365824
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000105854-PON2/tissue/adrenal+gland#imid_6365800
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105854-PON2/tissue/colon#imid_6365734
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105854-PON2/tissue/liver#imid_6365751
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PON2 IHC & IF
🧪Run appendix first: its glandular cells should stain (HPA: High in appendix glandular cells). Run salivary gland as a tissue negative (HPA: Not detected in salivary gland glandular cells); within the appendix slide, use cells without chromogen above background as internal references, without assuming a particular cell type is PON2-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PON2; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, a control immunoglobulin matched to the primary antibody’s host species, class and clonality, and a PON2 knockout sample or validated peptide-block control (standard IHC practice). Block endogenous peroxidase and check staining in appendix immune-cell areas against the no-primary control (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU M02027-1 tissue-IHC caption does not state a fixative (caption: fixative not stated). The caption uses heat retrieval in Tris-EDTA at pH 8.0–8.4 for 20 minutes, but does not establish that retrieval is required (caption: retrieval conditions). The supplied evidence does not establish whether frozen sections or IF are easier; in appendix, inspect immune-cell areas for peroxidase-related background (standard chromogenic IHC practice).
HPA tissue IHC evidence for PON2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Bergmann glia - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
Section 3
Advanced PON2 IHC Tips
Troubleshoot PON2 chromogenic IHC in paraffin sections with matched controls, documented processing, and cell level scoring.
What retrieval should I start with if PON2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in Tris-EDTA at pH 8.0 (datasheet M02027-1). The selected paraffin-section image used Tris-EDTA at pH 8.0–8.4 for 20 minutes, followed by 5% BSA blocking for 30 minutes (M02027-1 tissue-IHC caption). Keep heating time and section thickness consistent across cases, and compare weak sections with a concurrently processed positive control (standard IHC practice). If staining remains weak, test another retrieval buffer or heating time on serial sections while holding antibody concentration and detection constant (standard IHC practice). Score signal and tissue damage together, since excessive heating can compromise section morphology (standard IHC practice).
How should I assess whether fixation is suppressing PON2 staining?
PON2-specific fixation sensitivity is unknown from the supplied evidence; the selected image describes paraffin-embedded tissue but does not state its fixative (M02027-1 tissue-IHC caption). Record each specimen’s fixative and duration, then compare sections with matched processing and the same pH 8.0 Tris-EDTA retrieval (datasheet M02027-1; standard IHC practice). Include a concurrently processed positive control and inspect morphology before interpreting a weak result (standard IHC practice). If fixation histories differ, treat staining differences as provisional until matched controls show the assay behaves consistently (standard IHC practice). Do not infer fixation tolerance from tissue staining patterns or PON2 topology (HPA tissue IHC; UniProt Q15165 topology).
Does predominantly cytoplasmic PON2 staining fit its reported localisation?
A predominantly cytoplasmic pattern is compatible with the reported tissue-IHC profile, while cerebellar Bergmann glia have reported cytoplasmic or membrane staining (HPA tissue IHC). UniProt annotates PON2 as membrane associated without a transmembrane segment, so a sharply defined plasma membrane rim is not required to recognize plausible staining (UniProt Q15165 subcellular annotation and topology). Compare the compartment within intact cells against a no-primary control and against the same compartment in a positive reference section (standard IHC practice). HPA’s subcellular summary says “Secreted” but supplies no main location or ICC/IF images, so it does not resolve the tissue-level pattern (HPA subcellular). Avoid calling diffuse stain outside recognizable cells specific on localisation alone (standard IHC practice).
Could PON2 isoforms or processing alter what this antibody detects in sections?
PON2 has 3 listed isoforms and glycosylation sites at residues 254, 269, and 323 (UniProt Q15165 isoforms and glycosylation). The supplied evidence does not map the catalog antibody’s epitope, so isoform coverage and any effect of glycosylation on binding remain unknown (supplied M02027-1 evidence). Review any available immunogen or epitope documentation before assigning an unstained cell population to a particular isoform (standard antibody validation practice). Use the same retrieval and antibody conditions on serial sections when comparing cell populations, and report the antibody identity alongside the scoring method (standard IHC practice). An alternative antibody recognizing a documented, separate epitope can help investigate discordant staining if its tissue-IHC performance is established (standard antibody validation practice).
How can I investigate PON2 localisation by IF alongside chromogenic IHC?
Treat IF as a separate assay requiring its own antibody validation and controls; the supplied M02027-1 tissue image establishes paraffin-section IHC conditions, not IF performance (M02027-1 tissue-IHC caption). Multiplex PON2 with a validated marker for the cell population under examination, such as glia in a glial region, and inspect each channel separately (HPA tissue IHC; standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence and include single-stain and no-primary controls to assess bleed-through and background (standard IF practice). Because PON2 is membrane associated without a transmembrane segment and its antibody epitope is unmapped, determine epitope accessibility experimentally before selecting permeabilisation conditions (UniProt Q15165 topology; supplied M02027-1 evidence). Report fixation and permeabilisation separately for IF rather than transferring the paraffin IHC conditions (standard IF practice).
How do I distinguish PON2 signal from chromogenic background?
First compare the suspect deposit with a no-primary control on the same tissue and detection run (standard IHC practice). For peroxidase-based chromogenic detection, include an endogenous peroxidase block and check whether background persists before applying DAB; these are general IHC workflow steps, not PON2-specific findings (standard IHC practice). The selected image used 5% BSA for 30 minutes and primary antibody at 1:50 for 30 minutes at room temperature (M02027-1 tissue-IHC caption). If background is diffuse, compare a shorter primary incubation or greater dilution on adjacent sections while preserving the positive control (standard IHC practice). Assess pigment, section folds, edges, and necrotic areas microscopically before scoring cellular staining (standard IHC practice).
What should I quantify when comparing PON2 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since reported PON2 tissue staining is generally cytoplasmic and can include membrane staining in Bergmann glia (HPA tissue IHC). For interpretable chromogenic signal, record the percentage of positive target cells and intensity on a prespecified 0–3 scale, then calculate an H-score of 0–300 (standard IHC scoring practice). When cell abundance varies, report positive cells per mm² of viable tissue or within a defined cell compartment, with the denominator stated explicitly (standard quantitative IHC practice). Keep retrieval, detection, imaging, and threshold settings consistent, and exclude folds, section edges, and necrosis by predefined rules (standard quantitative IHC practice). Compare matched anatomical regions and retain a concurrently stained reference section to monitor run variation (standard IHC practice).
What would make an apparent PON2-positive region unconvincing?
Check whether stain lies in intact cells with a plausible cytoplasmic or membrane-associated pattern; a nuclear-only pattern needs independent validation (HPA tissue IHC; UniProt Q15165 subcellular annotation). A strong signal in expected populations, such as adrenal glandular cells or cerebral cortical glia, supports interpretation when controls behave as expected (HPA tissue IHC; standard IHC practice). Staining confined to a section edge, necrotic area, or no-primary control is more consistent with artefact than convincing cellular signal (standard IHC practice). For peroxidase detection, residual endogenous enzyme activity can produce misleading DAB deposits, so inspect the blocked control alongside the test slide (standard IHC practice). Because the HPA tissue profile has only medium staining–RNA consistency, avoid treating a single discrepant region as proof of absent or ectopic PON2 (HPA tissue IHC reliability description).
Boster reagents
Best PON2 / Serum paraoxonase/arylesterase 2 IHC Antibodies
M02027-1 has real IHC data from paraffin-embedded human colon carcinoma tissue (IHC image caption). Human, Mouse, and Rat reactivity is listed (catalog reactivity); IF data are absent (catalog IF images).
Real IHC data
Anti-PON2 Rabbit Monoclonal Antibody
Cat # M02027-1
M02027-1 is shown on paraffin-embedded human colon carcinoma tissue at 1:50 (IHC image caption). Its listed applications include IHC, and its listed reactivity covers Human, Mouse, and Rat (catalog applications/reactivity).
Which to pick: For tissue IHC, choose M02027-1, a rabbit monoclonal (catalog host/clone) shown on paraffin-embedded human colon carcinoma tissue (IHC image caption); the fixative is unreported (IHC image caption). For IF/ICC, no SKU in this payload has a listed IF application or IF image (catalog applications/IF images). For cross-species work, M02027-1 lists Human, Mouse, and Rat reactivity (catalog reactivity), while its IHC image documents human tissue only (IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15165 (PON2_HUMAN, Serum paraoxonase/arylesterase 2).
- Human Protein Atlas. PON2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PON2 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. PON2 antibody validation summary (2 antibodies).
- RHOA G17V induces T follicular helper cell specification and involves angioimmunoblastic T-cell lymphoma via upregulating the expression of PON2 through an NF-κB-dependent mechanism. Oncoimmunology 2022 — PMC9559328.
- Valproic acid inhibits glioblastoma multiforme cell growth via paraoxonase 2 expression. Oncotarget 2017 — PMC5362434.
- Paraoxonase 2 Facilitates Pancreatic Cancer Growth and Metastasis by Stimulating GLUT1-Mediated Glucose Transport. Molecular cell 2017 — PMC5567863.
- Role Played by Paraoxonase-2 Enzyme in Cell Viability, Proliferation and Sensitivity to Chemotherapy of Oral Squamous Cell Carcinoma Cell Lines. International journal of molecular sciences 2022 — PMC9820498.
- PubMed PMID:8661009 — UniProt-cited evidence.
- PubMed PMID:9714608 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PON2 (Serum paraoxonase/arylesterase 2). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC); membrane annotated (UniProt). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet M02027-1). Validated IHC antibody: M02027-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Esophagus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon carcinoma tissue; fixative not specified (datasheet M02027-1); 2. Fixation: Image fixative and duration unreported (datasheet M02027-1); 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 retrieval: Tris-EDTA pH 8.0 , 20 minutes (datasheet M02027-1); 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 monoclonal (clone 19P05) anti-PON2, 1:50 (datasheet M02027-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: A secreted variant may shift protein from RNA-defined sites (HPA tissue IHC); Isoform / epitope: 3 isoforms; verify which forms the epitope covers (UniProt); Regulation: Widely expressed; tissue levels vary (UniProt). Evidence: UniProt Q15165, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet M02027-1, PMC9559328, PMC5362434. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PON2/immunohistochemistry-ihc-guide.