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PELI2 / E3 ubiquitin-protein ligase pellino homolog 2 · IHC design guide
Design Immunohistochemistry for PELI2
Plan chromogenic PELI2 IHC in paraffin sections with catalog antibody A10176-1 (datasheet A10176-1). Use the nuclear and cytoplasmic tissue profile as a reference, while accounting for uncertain antibody specificity (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PELI2 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 | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | High respiratory epithelial staining; nuclear/cytoplasmic profile (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10176-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+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 | Antibody may detect more than one gene product (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one chain, residues 1–420 (UniProt) |
Section 1
Recommended PELI2 IHC & IF Protocols
The catalog antibody uses heat-mediated EDTA retrieval at pH 8.0 (datasheet A10176-1). The published mouse-section IHC excerpt reports detection and sample details (PMC7395429 methods).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A10176-1) |
| Fixation | Image fixative and duration unreported (datasheet A10176-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A10176-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10176-1) |
| Primary antibody | Rabbit anti-PELI2, 2-5 μg/ml (datasheet A10176-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10176-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10176-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PELI2-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0, as specified for this page’s catalog antibody (datasheet A10176-1).
Section 2
What Is the Expected PELI2 Staining Pattern?
In paraffin sections, expect nuclear and cytoplasmic staining in selected cells, including bronchial respiratory epithelium, skin keratinocytes and thyroid glandular cells (HPA tissue IHC: high; general nuclear and cytoplasmic profile). PELI2 has no transmembrane segment or signal peptide (UniProt Q9HAT8 topology and processing). Treat the pattern as provisional: HPA rates tissue IHC reliability Uncertain because the antibody targets proteins from more than one gene (HPA tissue IHC: Uncertain).
| Nuclear and cytoplasmic signal in bronchial respiratory epithelial cells or skin keratinocytes. | This matches reported high staining in those cells and the general compartment pattern (HPA tissue IHC: High; nuclear and cytoplasmic). Judge the result alongside a lower-staining comparator because the IHC assignment remains Uncertain (HPA tissue IHC: Uncertain). |
| A strong signal confined to membranes or extracellular material, without the reported nuclear or cytoplasmic pattern. | That distribution is discordant with the reported tissue pattern (HPA tissue IHC: general nuclear and cytoplasmic expression) and with the absence of a transmembrane segment or signal peptide (UniProt Q9HAT8 topology and processing). Check localization and background before calling it PELI2. |
| Strong staining in adipocytes, hippocampal glial cells or soft-tissue fibroblasts. | These cells were reported as not detected (HPA tissue IHC: Not detected). Consider cross-reactivity or endogenous detection activity (general IHC practice); a discrepant stain alone cannot establish which explanation applies, especially given the antibody-specific caution (HPA tissue IHC: Uncertain). |
| Diffuse chromogen across positive and comparator sections, obscuring cell boundaries. | The distribution cannot be scored confidently against HPA's cell-specific observations (HPA tissue IHC: cell-level staining). Review blocking, washes and detection controls for general background (general IHC practice) before interpreting apparent nuclear or cytoplasmic signal. |
| No signal in a well-preserved section containing bronchial respiratory epithelial cells. | That conflicts with the reported high signal in those cells (HPA tissue IHC: High). Check the IHC run and tissue identity first (general IHC practice); absence of staining does not by itself resolve the HPA antibody-specific uncertainty (HPA tissue IHC: Uncertain). |
💡Expected PELI2 appearanceA supportive positive result shows appreciable nuclear and cytoplasmic signal in reported high-staining cells such as bronchial respiratory epithelium (HPA tissue IHC: High; general nuclear and cytoplasmic profile); equally strong diffuse or membrane-only staining is suspect rather than confirmatory (HPA tissue IHC: Uncertain; UniProt Q9HAT8 topology).
| How strong is the tissue-pattern evidence? | HPA reports protein-level evidence and several high-staining cell populations, but rates this antibody's tissue IHC Uncertain because it targets proteins from more than one gene (HPA tissue IHC: protein evidence; Uncertain). A matching pattern is supportive, not proof of target specificity. |
| Which cells offer useful contrasts? | Bronchial respiratory epithelial cells and skin keratinocytes are reported High; bone-marrow hematopoietic cells are Medium; adipocytes and soft-tissue fibroblasts are Not detected (HPA tissue IHC). Compare identified cells within suitable sections rather than assigning one intensity to every cell in an organ. |
| Does PELI2 processing imply a surface or secreted pattern? | No signal peptide, propeptide or transmembrane segment is listed, and the recorded chain spans residues 1–420 (UniProt Q9HAT8 processing and topology). These annotations do not establish an IHC epitope, retrieval condition or fixation sensitivity. |
| What does IF/ICC add? | HPA reports approved nucleoplasm, nucleoli and basal-body localization, with additional cytosol and other structures, in ICC-IF (HPA subcellular: approved locations). That entry also cautions that its antibodies target proteins from multiple genes; it does not validate a paraffin-section compartment call (HPA subcellular: caution). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported high-staining cells show no chromogenic signal. | The run may have failed, or the chosen section may lack the specified cell population (general IHC practice; HPA tissue IHC: cell-level High calls). | Confirm tissue identity and preservation, inspect detection controls, and review the catalog antibody's IHC-P protocol for its stated conditions (general IHC practice). Avoid inventing a PELI2-specific retrieval requirement. |
| A high-staining control is positive, but adipocytes or fibroblasts stain just as strongly. | This disagrees with their Not detected entries (HPA tissue IHC). Cross-reactivity is plausible under the stated antibody caution; endogenous detection activity is another general IHC possibility. | Inspect no-primary and detection-only controls, then compare signal within the identified cell types (general IHC practice). Report the discordance and HPA's Uncertain reliability rather than scoring all staining as PELI2. |
| The stain appears exclusively membranous or extracellular. | The localization disagrees with the general nuclear and cytoplasmic tissue profile (HPA tissue IHC) and lacks support from PELI2 signal-peptide or transmembrane annotations (UniProt Q9HAT8). | Recheck morphology, chromogen deposits and control sections (general IHC practice). Record the observed compartment separately from the expected pattern; do not infer a PELI2 membrane pool from this stain alone. |
| Nuclear color is hard to distinguish from counterstain. | Heavy counterstaining or dense chromogen can obscure compartment scoring (general IHC practice); HPA describes both nuclear and cytoplasmic expression (HPA tissue IHC). | Compare the stained section with its controls and adjust counterstain or detection intensity using the local IHC workflow (general IHC practice). Re-score only cells whose compartments remain visible. |
| Patchy staining varies across a section. | Uneven reagent coverage or local background can create patchiness (general IHC practice); HPA's reported intensities refer to identified cells, not uniform staining throughout a tissue (HPA tissue IHC). | Check section quality, reagent coverage and washes, then compare the same cell type across interpretable areas (general IHC practice). Do not assign a tissue-wide negative call from an area lacking the relevant cells. |
| Nucleoli or basal bodies appear positive in an IF image, but the paraffin-section stain looks different. | Those approved locations come from ICC-IF, while tissue IHC is summarized more broadly as nuclear and cytoplasmic (HPA subcellular; HPA tissue IHC). The ICC-IF entry carries its own multiple-gene antibody caution (HPA subcellular). | Score the paraffin section against the tissue IHC profile and its reliability note (HPA tissue IHC: Uncertain). Treat the ICC-IF image as context, without requiring punctate structures to be visible by chromogenic IHC. |
Experiment evidence
IHC & IF images showing PELI2 staining
A10176-1 · IHCIHC analysis of PELI2 using anti-PELI2 antibody (A10176-1). PELI2 was detected in a paraffin-embedded section of human colorectal adenocarci
A10176-1 · IHCIHC analysis of PELI2 using anti-PELI2 antibody (A10176-1). PELI2 was detected in a paraffin-embedded section of human lung adenocarcinoma t
A10176-1 · IFIF analysis of PELI2 using anti-PELI2 antibody (A10176-1). PELI2 was detected in an immunocytochemical section of HELA cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000139946-PELI2/tissue/cerebral+cortex#imid_16195743
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000139946-PELI2/tissue/colon#imid_16189349
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000139946-PELI2/tissue/liver#imid_16189350
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000139946-PELI2/tissue/kidney#imid_16189351
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA053182; green = PELI2, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA053182; green = PELI2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PELI2 IHC & IF
🧪Run bronchus first and expect staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells); use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes). On the bronchus slide, use cells without signal above the no-primary background as internal negatives; the supplied HPA rows do not identify a specific negative cell type within bronchus (HPA: bronchus row).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PELI2 in CACO-2, HeLa, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (approved), Nucleoli (approved), Basal body (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and matched rabbit IgG isotype controls, plus PELI2 knockout material or a validated peptide-block control where available (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check whether peroxidase-positive inflammatory cells in bronchus produce background DAB staining (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative used for the selected A10176-1 tissue-IHC image is unreported (selected-SKU caption). The caption documents heat-mediated EDTA retrieval at pH 8.0 for a paraffin section, but does not establish that retrieval is required for PELI2 (selected-SKU caption). Paraffin IHC has a documented example, while HPA lists ICC-IF images without a matched frozen-section comparison, so whether frozen sections or IF are easier is unreported (selected-SKU caption; HPA: subcellular record); endogenous peroxidase in bronchial inflammatory cells can complicate DAB interpretation (standard chromogenic IHC practice).
HPA tissue IHC evidence for PELI2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Caution, targets protein from more than one gene. Pending external verification.). 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Skin | Keratinocytes | High | Protein (IHC) | HPA → |
Section 3
Advanced PELI2 IHC Tips
Troubleshoot PELI2 staining in paraffin sections by checking retrieval, controls, compartment, and cell identity before interpreting DAB signal.
Which retrieval conditions should I start with for PELI2 IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 for PELI2 paraffin-section IHC (datasheet A10176-1). The demonstrated section was then stained with 2 μg/ml catalog antibody overnight at 4°C, so keep those conditions consistent while assessing retrieval (datasheet A10176-1). The caption gives no heating duration; record the instrument, temperature, heating time, and cooling time for each run (datasheet A10176-1; standard IHC practice). If staining is weak, adjust heating time on matched sections before testing another buffer, using a positive control and a no-primary control to distinguish recovery from nonspecific staining (standard IHC practice).
Could fixation explain weak or uneven PELI2 staining?
The selected image documents a paraffin-embedded section but does not report its fixative, so target-specific fixation sensitivity is unknown (datasheet A10176-1). Record the actual fixative, fixation duration, tissue thickness, and processing history before comparing sections, because those variables can change antigen accessibility in IHC (standard IHC practice). Compare suspect and reference sections in the same staining run with EDTA retrieval at pH 8.0 and the same antibody concentration (datasheet A10176-1; standard IHC practice). If signal differs, test matched tissue processed under documented conditions; do not attribute the difference to a PELI2-specific fixation effect without controlled evidence (standard IHC practice).
How should I assess nuclear versus cytoplasmic PELI2 staining?
Score nuclear and cytoplasmic DAB staining separately: tissue IHC reports a general nuclear and cytoplasmic pattern, but its reliability is uncertain because the antibodies may detect proteins from multiple genes (HPA tissue IHC). UniProt provides no subcellular annotation for PELI2, so compartment alone cannot establish specificity (UniProt Q9HAT8). The HPA cell imaging summary includes nucleoplasm, nucleoli, basal body, and additional cytosolic locations, with the same cross-reactivity caution (HPA subcellular). Compare compartment patterns in morphologically intact cells across a positive control and matched no-primary section; investigate diffuse deposits or staining confined to damaged edges before scoring (standard IHC practice).
What if the antibody stains only some cell compartments?
UniProt lists a single 1–420 PELI2 chain and no annotated isoforms, while the supplied antibody evidence does not identify its epitope (UniProt Q9HAT8; datasheet A10176-1). PELI2 has an FHA domain spanning residues 15–202, but the antibody cannot be assigned to that region from these data (UniProt Q9HAT8; datasheet A10176-1). If compartments differ between sections, check retrieval and processing records before proposing epitope masking (standard IHC practice). Compare staining with an independently validated antibody recognizing a documented, distinct epitope when available, and interpret agreement alongside cell identity and controls rather than assuming an unreported isoform (standard IHC practice; UniProt Q9HAT8).
How can IF help check the cell type and compartment?
For a separate IF experiment, multiplex PELI2 with an epithelial marker such as pan-cytokeratin when examining respiratory epithelium, which HPA reports as strongly stained in bronchus (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore after checking unstained tissue for autofluorescence, and include single-label controls to assess channel bleed-through (standard IF practice). Permeabilise for intracellular epitopes: PELI2 has no annotated transmembrane segment, and HPA imaging reports nuclear and cytosolic locations (UniProt Q9HAT8; HPA subcellular). Titrate the IF antibody and permeabilisation conditions independently; the supplied paraffin-section caption establishes only an IHC workflow for A10176-1 (datasheet A10176-1).
How do I separate PELI2 signal from DAB background?
The demonstrated IHC section used 10% goat serum before 2 μg/ml rabbit antibody overnight at 4°C (datasheet A10176-1). Begin troubleshooting by keeping that blocking and primary-antibody sequence fixed while comparing a no-primary section with the stained section (datasheet A10176-1; standard IHC practice). Include a peroxidase block before HRP and DAB development, and check whether deposits persist without primary antibody; those are general chromogenic-IHC controls, not evidence specific to PELI2 (standard IHC practice). If background remains, inspect wash quality, section edges, necrotic areas, and DAB development time before changing antibody concentration (standard IHC practice).
What is a defensible way to quantify PELI2 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear or cytoplasmic compartment before scoring, since HPA describes both compartments in tissue IHC (HPA tissue IHC; standard IHC practice). Within that population, report the percentage of positive cells and an H-score on a 0–300 scale, or report positive-cell density per mm² if cell counts suit the question (standard IHC practice). Normalise counts to the number of eligible cells or measured viable tissue area, and apply identical thresholds across sections stained in one run (standard IHC practice). Record excluded edges and necrosis, and report control performance because HPA rates its tissue antibody evidence uncertain (HPA tissue IHC; standard IHC practice).
When is a PELI2-positive DAB pattern convincing?
A convincing result shows reproducible staining in intact, identifiable cells and little signal in a matched no-primary section (standard IHC practice). Respiratory epithelial cells in bronchus and glandular cells in thyroid are reported as high, whereas adipocytes are reported as not detected; these are comparison patterns, not definitive specificity controls (HPA tissue IHC). Examine apparent nuclear or cytoplasmic signal against morphology, because HPA reports both compartments while warning that its antibodies may recognize proteins from multiple genes (HPA tissue IHC). Treat edge-only deposits, necrotic staining, and signal remaining after primary omission as artefact candidates, including possible endogenous peroxidase activity (standard IHC practice).
Boster reagents
Best PELI2 / E3 ubiquitin-protein ligase pellino homolog 2 IHC Antibodies
A10176-1 has human paraffin-section IHC images and a HeLa-cell IF image (catalog image captions); its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-PELI2 Antibody ®
Cat # A10176-1
A10176-1 has IHC images from paraffin sections of human colorectal adenocarcinoma, lung adenocarcinoma and tonsil (A10176-1 IHC captions). The same SKU has an IF image from HeLa cells (A10176-1 IF caption).
Which to pick: Choose A10176-1 for human tissue IHC: its paraffin-section captions use 2 μg/ml antibody after heat retrieval in EDTA, pH 8.0 (A10176-1 IHC captions); the fixative is unreported (A10176-1 IHC captions). For IF/ICC, A10176-1 is listed for both applications and has a HeLa-cell IF image at 5 μg/ml (catalog applications; A10176-1 IF caption). For mouse or rat work, A10176-1 lists reactivity with both species, but the supplied IHC and IF images show human samples only; clonality is unspecified (catalog reactivity; A10176-1 image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9HAT8 (PELI2_HUMAN, E3 ubiquitin-protein ligase pellino homolog 2).
- Human Protein Atlas. PELI2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. PELI2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm, nucleoli and basal body. In addition localized to the cytosol, intermediate filaments, centrosome and centriolar satellites. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PELI2 antibody validation summary (1 antibodies).
- PELI2 inhibits colorectal cancer development through MAPK signaling pathway. Molecular medicine (Cambridge, Mass.) 2025 — PMC12160117.
- Dibutyl phthalate promotes juvenile Sertoli cell proliferation by decreasing the levels of the E3 ubiquitin ligase Pellino 2. Environmental health : a global access science source 2020 — PMC7395429.
- PubMed PMID:11306823 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PELI2 (E3 ubiquitin-protein ligase pellino homolog 2). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10176-1). Validated IHC antibody: A10176-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A10176-1); 2. Fixation: Image fixative and duration unreported (datasheet A10176-1); 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: EDTA pH 8.0 (datasheet A10176-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10176-1); 8. Primary antibody: Rabbit anti-PELI2, 2-5 μg/ml (datasheet A10176-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10176-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody may detect more than one gene product (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one chain, residues 1–420 (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q9HAT8, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm, nucleoli and basal body. In…, Datasheet A10176-1, PMC7395429. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PELI2/immunohistochemistry-ihc-guide.