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PEBP1 / Phosphatidylethanolamine-binding protein 1 · IHC design guide
Design Immunohistochemistry for PEBP1
Plan PEBP1 paraffin-section IHC using liver hepatocytes or kidney proximal tubules as high-staining reference cells (HPA tissue IHC). The guide covers cytoplasmic scoring (HPA tissue IHC) and the catalog antibody’s chromogenic workflow (datasheet A01668).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PEBP1 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 | Cytoplasm (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues; high in hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01668) | |
| Positive control | Adrenal gland+4 more · see all | |
| 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01668) | |
| Caveat | Endogenous peroxidase may mimic liver DAB staining (standard IHC practice) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–187 (UniProt) |
Section 1
Recommended PEBP1 IHC & IF Protocols
The catalog antibody protocol uses citrate pH 6 retrieval (datasheet A01668). Two published PEBP1 chromogenic IHC protocols provide tissue specific examples (PMC13067198; PMC11177062).
| Sample | Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01668) |
| Fixation | Image fixative and duration unreported (datasheet A01668); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01668) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01668) |
| Primary antibody | Rabbit anti-PEBP1, 0.5-1μg/ml (datasheet A01668) |
| Primary incubation | Overnight at 4 °C (datasheet A01668) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01668) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PEBP1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a majority of tissues. Highest levels in liver, kidney, endocrine tissues and brain. No signal in the no-primary control. |
💡Decision noteStart with heat mediated citrate pH 6 retrieval: it matches the catalog protocol and both published methods (datasheet A01668; PMC13067198; PMC11177062).
Section 2
What Is the Expected PEBP1 Staining Pattern?
In paraffin sections, expect mainly cytoplasmic PEBP1 staining: UniProt places PEBP1 in the cytoplasm and reports no transmembrane segment (UniProt P30086 topology). HPA reports cytoplasmic expression in most tissues, with high staining in hepatocytes, kidney proximal tubules and several endocrine cell populations (HPA tissue IHC). Its tissue IHC profile has Enhanced reliability, reflecting high consistency between staining and RNA expression (HPA tissue IHC).
| Clear cytoplasmic staining in hepatocytes or kidney proximal tubule cell bodies. | This fits high PEBP1 staining in these cell populations (HPA: High in hepatocytes and proximal tubules) and its reported cytoplasmic location (UniProt P30086). Judge the cellular pattern alongside controls, since stain intensity alone does not establish antibody specificity (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear dominant IHC pattern conflicts with the reported cytoplasmic tissue pattern (HPA tissue IHC; UniProt P30086). Treat it as a possible artefact and check controls, staining distribution and detection conditions before assigning it to PEBP1 (general IHC practice). |
| Strong staining in adipocytes, germinal center cells or smooth muscle cells. | These specific cell populations were not detected in the HPA tissue IHC record; that finding does not make their entire tissues negative (HPA: Not detected in adipocytes, germinal center cells and smooth muscle cells). Unexpected staining warrants checks for cross-reactivity or endogenous detection activity (general IHC practice). |
| A broad haze covers cells and surrounding tissue without clear cellular boundaries. | Diffuse background cannot be read as the reported cytoplasmic cell pattern (HPA tissue IHC). Assess the no-primary control, blocking, washes and chromogen development to identify nonspecific reagent binding or detection background (general IHC practice). |
| No staining in hepatocytes or kidney proximal tubules on a suitable section. | That conflicts with HPA's high staining in those cell populations (HPA tissue IHC), but a single absent signal does not prove PEBP1 is absent. Check section quality, retrieval, primary antibody conditions and detection controls before interpreting the sample (general IHC practice). |
💡Expected PEBP1 appearanceCall a convincing positive result when cytoplasmic stain is clear and relatively strong in hepatocytes or kidney proximal tubule cell bodies (HPA: High; UniProt P30086 cytoplasm); isolated nuclear stain or uniform tissue haze is suspect (general IHC practice).
| Cellular compartment | PEBP1 is reported in the cytoplasm, with no transmembrane segment or signal peptide (UniProt P30086). For tissue IHC, use the observed cytoplasmic pattern as the main localization reference (HPA tissue IHC). |
| Cell type and tissue context | High staining is reported in adrenal, parathyroid and thyroid glandular cells and testis Leydig cells; breast glandular cells and caudate neurons are Medium (HPA tissue IHC). Compare the named cell population, rather than treating every cell in a tissue as equally positive (general IHC practice). |
| Strength of tissue evidence | The overall tissue profile has Enhanced reliability from agreement between staining and RNA expression (HPA tissue IHC). Individual antibodies HPA008819 and CAB013493 have Enhanced IHC validation, while CAB080063 is Supported (HPA antibodies); these labels do not validate an unlisted catalog antibody. |
| IF/ICC Q: where is PEBP1 seen? | A: HPA reports approved cytosol and plasma membrane localization in ICC-IF (HPA subcellular). That observation provides context for the separate IF/ICC guide; tissue IHC interpretation follows the cytoplasmic HPA tissue profile (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| High positive control is blank. | Primary antibody, retrieval or detection may have failed; the expected high cell populations are hepatocytes and kidney proximal tubules (HPA tissue IHC; general IHC practice). | Confirm that the named cells are present and intact, then check the antibody's IHC-P instructions, retrieval step and detection reagents with a positive control (general IHC practice). |
| Signal is mainly nuclear. | The dominant compartment disagrees with cytoplasmic tissue staining (HPA tissue IHC; UniProt P30086); nonspecific staining or a detection artefact is possible (general IHC practice). | Review the no-primary control and staining distribution, and verify the primary antibody and detection setup before scoring nuclei as positive (general IHC practice). |
| Expected negative cell populations stain strongly. | HPA reports no detection in adipocytes, germinal center cells and smooth muscle cells (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Identify the stained cell type, inspect no-primary controls and apply the appropriate endogenous activity block for the detection system (general IHC practice). |
| The whole section has diffuse brown background. | Nonspecific binding, inadequate washing or excessive chromogen development can obscure cellular staining (general IHC practice). | Compare no-primary and positive controls; review blocking, wash steps and development time before judging PEBP1 localization (general IHC practice). |
| Signal is weak in a known high cell population. | Assay conditions may be limiting detection, although HPA reports high staining in hepatocytes and proximal tubules (HPA tissue IHC; general IHC practice). | Check section quality and antibody instructions, then optimize retrieval and primary antibody conditions using the same positive control (general IHC practice). No PEBP1-specific retrieval response is supplied. |
| A low or negative HPA cell population has little signal. | That can agree with the cell-specific reference pattern; HPA reports Low in bronchial ciliated cell bodies and Not detected in esophageal squamous epithelial cells (HPA tissue IHC). | Verify signal in a high reference cell population before calling an assay failure, and record the exact cell type assessed (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PEBP1 staining
A01668 · IHCIHC analysis of PBP using anti-PBP antibody (A01668). PBP was detected in paraffin-embedded section of human intestinal cancer tissue. Heat
A01668 · IHCIHC analysis of PBP using anti-PBP antibody (A01668). PBP was detected in paraffin-embedded section of human sarcoma tissue. Heat mediated a
M01668 · IHCImmunohistochemical analysis of paraffin-embedded human liver, using PEBP1 Antibody.
M01668 · IFImmunofluorescent analysis of Hela cells, using PEBP1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000089220-PEBP1/tissue/cerebral+cortex#imid_3558336
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000089220-PEBP1/tissue/adrenal+gland#imid_3558312
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000089220-PEBP1/tissue/colon#imid_3547462
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000089220-PEBP1/tissue/liver#imid_3547471
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA063904; green = PEBP1, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA063904; green = PEBP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PEBP1 IHC & IF
🧪Run kidney first; proximal tubule cell bodies should stain (HPA: High in kidney proximal tubules). Run adipose tissue as the negative, scoring adipocytes for absent staining (HPA: Not detected in adipocytes); on the kidney slide, cells without chromogen can show background levels, but the supplied HPA rows do not identify a specific renal cell type as an internal negative (HPA: kidney proximal tubules High).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PEBP1 in Hep-G2, U2OS, A-431, U-251MG, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, and process PEBP1 knockout material alongside as a biological negative if available (caption: rabbit primary antibody; standard IHC practice). In kidney sections, quench endogenous peroxidase and check for endogenous biotin before interpreting signal from the biotin/SABC-DAB detection system (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01668 tissue-IHC caption does not state the fixative (caption: fixative not stated). Its paraffin-section workflow uses heat retrieval in citrate buffer at pH 6 for 20 minutes; use that documented condition as a starting point and optimize retrieval empirically (caption: citrate retrieval). The paraffin IHC workflow is documented, while the supplied evidence does not establish that frozen sections or IF are easier; kidney proximal tubules also require attention to endogenous biotin when using SABC detection (caption: paraffin section and SABC; HPA: High in kidney proximal tubules; standard IHC practice).
HPA tissue IHC evidence for PEBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Proximal tubules (cell body) | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Basal cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PEBP1 IHC Tips
Troubleshoot PEBP1 staining in paraffin section IHC using the catalog antibody’s tissue image and independent localisation evidence.
How should I adjust retrieval when PEBP1 staining is weak?
Start with heat mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01668; A01668 tissue IHC caption). Keep section thickness, heating method and primary incubation constant while comparing retrieval runs (standard IHC practice). The catalog image used 1 μg/mL primary antibody overnight at 4°C, so match that incubation before attributing weak staining to retrieval (A01668 tissue IHC caption). If staining remains weak, test a modest change in heating duration on adjacent sections and inspect tissue integrity (standard IHC practice). Use hepatocytes or kidney proximal tubules as positive reference cells when available (HPA tissue IHC: High in both).
Could fixation explain weak or uneven PEBP1 staining?
Target specific fixation sensitivity is unknown: the selected tissue IHC caption identifies a paraffin embedded section but does not state its fixative (A01668 tissue IHC caption). Record the fixative, fixation duration and processing history for each specimen before comparing staining intensity (standard IHC practice). Compare matched sections processed together, and check whether weak regions coincide with poor morphology or uneven reagent access (standard IHC practice). Keep the established citrate pH 6, 20 minute retrieval condition fixed during that comparison (datasheet A01668; A01668 tissue IHC caption). Do not use tissue expression patterns to assign a fixation effect without a controlled fixation comparison (standard IHC practice).
Which staining compartment should count as PEBP1 positive?
Score cytoplasmic staining in the relevant cells first, because tissue IHC shows cytoplasmic expression in a majority of tissues (HPA tissue IHC). Cytosolic staining also agrees with the protein’s annotated location, while plasma membrane staining has independent cellular imaging support (UniProt P30086 subcellular; HPA subcellular: Cytosol and Plasma membrane approved). PEBP1 has no annotated transmembrane segment, so membrane staining alone needs careful corroboration (UniProt P30086 topology; standard IHC practice). Review membrane accentuation against adjacent cytoplasm and cell boundaries at the same exposure and counterstain level (standard IHC practice). Predominantly nuclear signal should prompt control review before it enters a PEBP1 score (UniProt P30086 subcellular; standard IHC practice).
Could an isoform or modified epitope change the IHC pattern?
The supplied record lists 0 isoforms and a PEBP1 chain spanning residues 2–187 (UniProt P30086 isoforms and processing). It also lists phosphorylation sites, including serines 6 and 13, but provides no epitope map for the catalog antibody (UniProt P30086 modified residues; A01668 evidence supplied). Therefore, a changed staining pattern cannot be assigned to isoform selection or phosphorylation dependent recognition from these data alone (standard IHC interpretation). Compare the catalog antibody with an independently validated antibody recognizing a documented, distinct epitope if epitope dependence matters (standard IHC practice). Keep retrieval and detection conditions matched while assessing agreement between the two reagents (standard IHC practice).
How can IF help resolve ambiguous PEBP1 localisation?
Use IF/ICC as a separate localisation experiment and multiplex PEBP1 with a marker that identifies the expected cell type in the specimen (standard IF practice). For example, hepatocytes or kidney proximal tubules are high expressing IHC reference cells, but the companion marker must be chosen and validated for the specimen (HPA tissue IHC: High in both; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, and keep emission channels sufficiently separated for reliable colocalisation (standard IF practice). Set permeabilisation according to the mapped epitope’s side of the membrane; the supplied evidence does not locate the catalog antibody’s epitope (standard IF practice; A01668 evidence supplied). Check cytosol and cell boundaries separately because both have localisation support (HPA subcellular: Cytosol and Plasma membrane approved).
What should I check when DAB staining is widespread?
Compare a no primary control with the complete stain to locate background arising from detection reagents (standard IHC practice). The catalog tissue image used 10% goat serum, a biotinylated secondary, an avidin based complex and DAB, so evaluate blocking and endogenous biotin when reproducing that workflow (A01668 tissue IHC caption; standard IHC practice). Include an endogenous peroxidase block before DAB development, then inspect whether residual colour persists without primary antibody (standard IHC practice). Titrate primary antibody around the image’s 1 μg/mL condition and keep development time consistent across sections (A01668 tissue IHC caption; standard IHC practice). Judge background against cell morphology and the expected cytoplasmic pattern (HPA tissue IHC; standard IHC practice).
How should I quantify PEBP1 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, using morphology and a consistent region selection rule across specimens (standard IHC practice). For cytoplasmic PEBP1, record the percentage of cells at each intensity level and calculate an H-score on a 0–300 scale (HPA tissue IHC: Cytoplasmic expression; standard IHC scoring). Report percentage positive cells separately, with one positivity threshold applied to all comparable slides (standard IHC practice). If counting discrete positive cells, report density per mm² of viable tissue and exclude necrotic or missing areas (standard IHC practice). Normalise comparisons to eligible cells or viable tissue area, and analyse batches with the same retrieval and DAB development conditions (standard IHC practice).
How do I distinguish true PEBP1 staining from artefact?
A convincing result identifies the stained cell type and shows cytoplasmic signal consistent with the tissue IHC profile (HPA tissue IHC). Strong staining in hepatocytes or kidney proximal tubules provides a useful reference, while absent staining in adipocytes or lymph node germinal center cells can inform a comparison without serving as an absolute negative control (HPA tissue IHC: High and Not detected, respectively; standard IHC interpretation). Review predominantly nuclear colour, section edge accentuation and necrotic regions as potential artefacts (UniProt P30086 subcellular; standard IHC practice). Run a no primary control to assess endogenous enzyme or detection reagent colour (standard IHC practice). Accept membrane accentuation only when its cell boundaries and controls support it (HPA subcellular: Plasma membrane approved; standard IHC practice).
Boster reagents
Best PEBP1 / Phosphatidylethanolamine-binding protein 1 IHC Antibodies
Both antibodies have human paraffin-section IHC images (IHC captions); M01668 also has a HeLa IF image (IF caption). Listed reactivity includes human and mouse for both, and rat for M01668 (catalog reactivity).
Real IHC data
Anti-PBP/PEBP1 Antibody ®
Cat # A01668
Real IHC data
Anti-PEBP1/Pbp Rabbit Monoclonal Antibody
Cat # M01668
A01668 will render with an IHC image from a paraffin-embedded human intestinal cancer section (A01668 IHC caption); its listed reactivity is human and mouse (catalog reactivity). M01668 will render with an IHC image from paraffin-embedded human liver (M01668 IHC caption); it is also listed for ICC/IF and has a HeLa IF image (catalog applications; M01668 IF caption).
Which to pick: For tissue IHC, choose A01668 if its rabbit polyclonal format and human intestinal cancer paraffin-section example fit the experiment (catalog host/clonality; A01668 IHC caption). For IF/ICC or rat samples, choose M01668: it is a rabbit monoclonal listed for ICC/IF and human, mouse and rat reactivity, with a HeLa IF image and a human liver paraffin-section IHC image (catalog host/clonality, applications and reactivity; M01668 IF and IHC captions). The fixative is unreported for both paraffin-section IHC examples (A01668 and M01668 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30086 (PEBP1_HUMAN, Phosphatidylethanolamine-binding protein 1).
- Human Protein Atlas. PEBP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PEBP1 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. PEBP1 antibody validation summary (5 antibodies).
- Expression Analysis of Selected microRNAs in Diabetes Mellitus Visceral Fat Tissues. Journal of obesity 2026 — PMC13067198.
- RIPK4/PEBP1 axis promotes pancreatic cancer cell migration and invasion by activating RAF1/MEK/ERK signaling. International journal of oncology 2018 — PMC5843398.
- Metabolomics reveals that ferroptosis participates in bisphenol A-induced testicular injury. Heliyon 2024 — PMC11177062.
- Drosophila PEBP1 inhibits intestinal stem cell aging via suppression of ERK pathway. Oncotarget 2018 — PMC5915051.
- PubMed PMID:8144042 — UniProt-cited evidence.
- PubMed PMID:7637590 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PEBP1 (Phosphatidylethanolamine-binding protein 1). Expected localisation Cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01668). Validated IHC antibody: A01668 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01668); 2. Fixation: Image fixative and duration unreported (datasheet A01668); 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: Citrate pH 6 , 20 min (datasheet A01668); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01668); 8. Primary antibody: Rabbit anti-PEBP1, 0.5-1μg/ml (datasheet A01668); 9. Primary incubation: Overnight at 4 °C (datasheet A01668). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01668); Caveat: Endogenous peroxidase may mimic liver DAB staining (standard IHC practice); Isoform / epitope: No annotated isoforms; mature chain spans residues 2–187 (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt P30086, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and cytosol., Datasheet A01668, PMC13067198, PMC11177062. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PEBP1/immunohistochemistry-ihc-guide.