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NPTXR / Neuronal pentraxin receptor · IHC design guide
Design Immunohistochemistry for NPTXR
Plan NPTXR chromogenic IHC on paraffin sections with the catalog antibody's 2–5 μg/ml range (datasheet A15505-1). Use cytoplasmic staining in a subset of cortical neurons as the tissue reference, while accounting for its uneven distribution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NPTXR 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 in a subset of cortical neurons (HPA tissue IHC) | |
| Staining pattern | A subset of cortical neurons shows cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A15505-1) | |
| Positive control | Cerebral cortex | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Only a subset of cortical neurons stains (HPA tissue IHC) | |
| Regulation | Brain-enriched RNA expression (HPA RNA) | |
| Isoform / epitope | No isoforms; extracellular 24–500 vs cytoplasmic 1–2 matters (UniProt) |
Section 1
Recommended NPTXR IHC & IF Protocols
This section pairs the catalog antibody’s IHC-P protocol (datasheet A15505-1) with a published gastric cancer IHC method (PMC7448342).
| Sample | Paraffin-embedded human glioma tissue; fixative not specified (datasheet A15505-1) |
| Fixation | Image fixative and duration unreported (datasheet A15505-1); 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 A15505-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A15505-1) |
| Primary antibody | Rabbit anti-NPTXR, 2-5 μg/ml (datasheet A15505-1) |
| Primary incubation | Overnight at 4 °C (datasheet A15505-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A15505-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NPTXR-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a subset of neuronal cells in cerebral cortex. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A15505-1); the published method does not specify retrieval conditions (PMC7448342).
Section 2
What Is the Expected NPTXR Staining Pattern?
In paraffin-section IHC, expect cytoplasmic staining in a subset of cerebral cortex neuronal cells, with medium intensity in HPA's observed pattern (HPA tissue IHC: Enhanced; external verification pending). NPTXR is a membrane protein with a transmembrane segment at residues 3–23 and an extracellular region at 24–500 (UniProt O95502 topology). Membrane-associated signal can therefore be plausible, but the cortical IHC reference pattern is cytoplasmic (HPA tissue IHC).
| A subset of cortical neuronal cells shows medium cytoplasmic chromogen, while neighboring cells stain less or remain clear. | This matches the observed positive reference: HPA reports medium cytoplasmic expression in a subset of cerebral cortex neuronal cells (HPA tissue IHC). Judge the pattern cell by cell; the reference does not imply that every cortical cell should stain. |
| Signal follows neuronal cell outlines as well as appearing in the cytoplasm. | A membrane component is compatible with NPTXR topology (UniProt O95502 topology) and approved plasma membrane localization in ICC-IF (HPA subcellular). For chromogenic IHC, compare the full pattern with HPA's observed cytoplasmic neuronal staining before calling a result concordant (HPA tissue IHC). |
| Strong nuclear-only staining dominates the cortical section. | This differs from the reported cortical tissue-IHC pattern and calls for control review (HPA tissue IHC). It is not proof of an artefact: HPA also reports nucleoplasmic localization as an additional ICC-IF location (HPA subcellular). Assess compartment, cell identity, and background together. |
| Broad staining appears in cells outside the expected cortical neuronal subset, including cells in an HPA negative reference. | Treat this as possible cross-reactivity or endogenous detection activity, then inspect negative controls (general IHC practice). HPA reports no detection in adipocytes of adipose tissue and glial cells of caudate; those are cell-specific observations, not whole-tissue guarantees (HPA tissue IHC). |
| The known-positive cortical neuronal subset has no discernible signal. | A blank result conflicts with the HPA cortical reference, but alone does not establish biological absence (HPA tissue IHC; general IHC practice). Check section quality, controls, retrieval, antibody and detection setup before interpreting a negative result. HPA reports low staining in hippocampal neuronal cells, making that a weaker positive reference (HPA tissue IHC). |
💡Expected NPTXR appearanceCall the result positive when a subset of cerebral cortex neuronal cells shows predominantly cytoplasmic, medium-intensity staining (HPA tissue IHC); widespread staining across unrelated cells or uniform background is suspect (general IHC practice).
| Tissue and cell selection | Cerebral cortex neuronal cells provide the medium-staining reference; hippocampal neuronal cells are reported at low level (HPA tissue IHC). Use the cortical subset to assess sensitivity, and interpret a weak hippocampal result against its lower reported level. |
| Membrane topology | NPTXR has a transmembrane segment at residues 3–23 and an extracellular region at 24–500 (UniProt O95502 topology). Epitope position is not supplied, so topology cannot specify antigen retrieval conditions or predict whether this antibody will emphasize the membrane. |
| Protein processing and glycosylation | UniProt lists one chain spanning residues 1–500, no annotated signal peptide or propeptide, and glycosylation sites at 42, 216, and 463 (UniProt O95502). These annotations do not establish shedding, fixation sensitivity, or an optimal retrieval method. |
| Antibody validation | The supplied HPA antibody has Enhanced IHC status, and HPA rates the tissue staining Enhanced for consistency with RNA expression while external verification remains pending (HPA antibody; HPA tissue IHC). These ratings support use of the reference pattern; they do not validate every section or detection run. |
| IF/ICC Q: Where should signal localize? | A: HPA approves plasma membrane and cytosol as main ICC-IF locations and nucleoplasm as an additional location (HPA subcellular). That ICC-IF record provides a localization comparison, not an IF/ICC protocol or a reason to replace the cortical tissue-IHC reference. |
| Situation | Likely cause | Next action |
|---|---|---|
| Cortical neuronal staining is absent. | The result conflicts with the medium cortical reference; the failed step cannot be identified from appearance alone (HPA tissue IHC; general IHC practice). | Review a positive control, section integrity, retrieval, antibody application, and chromogenic detection in sequence (general IHC practice). Avoid declaring NPTXR absent until the run controls work. |
| Cortical neurons stain weakly. | Low assay signal or sampling of cells outside the reported positive subset may explain the result (HPA tissue IHC; general IHC practice). No NPTXR-specific fixation effect is established by the supplied sources. | Compare several intact neuronal cells with a successful control and optimize retrieval or antibody conditions using the assay instructions (general IHC practice). Do not infer a target-specific retrieval setting from UniProt topology. |
| Diffuse chromogen obscures cell boundaries. | Background from blocking, washing, antibody conditions, or detection can mask the subset pattern (general IHC practice). Diffuse staining is unlike HPA's reported cortical neuronal pattern (HPA tissue IHC). | Inspect the negative reagent control, then review blocking, wash steps, antibody concentration, and detection time (general IHC practice). Score NPTXR only after individual cells and compartments can be resolved. |
| An HPA negative reference cell population stains strongly. | Possible nonspecific binding or endogenous detection activity should be considered (general IHC practice). HPA reports no detection in adipose adipocytes and caudate glial cells (HPA tissue IHC). | Check cell identity and the negative reagent control; review blocking and detection chemistry before assigning target positivity (general IHC practice). Compare the specified cell population, rather than treating its entire tissue as negative. |
| Only nuclei stain in the cortical section. | Nuclear-only IHC diverges from HPA's cytoplasmic cortical pattern, although HPA lists nucleoplasm as an additional ICC-IF location (HPA tissue IHC; HPA subcellular). | Recheck the positive control, background, and neuronal cell identity (general IHC practice). Record the nuclear pattern as discordant with the tissue-IHC reference unless independent evidence supports it. |
| All cortical cells stain with similar intensity. | Uniform staining conflicts with the reported positive subset and may reflect excess background or nonspecific detection (HPA tissue IHC; general IHC practice). | Compare stained and unstained cell populations, inspect the negative reagent control, and adjust assay conditions if background is present (general IHC practice). Reassess the pattern against HPA's medium neuronal-subset reference. |
Experiment evidence
IHC & IF images showing NPTXR staining
A15505-1 · IHCIHC analysis of NPTXR using anti-NPTXR antibody (A15505-1). NPTXR was detected in a paraffin-embedded section of human glioma tissue. Heat m
A15505-1 · IHCIHC analysis of NPTXR using anti-NPTXR antibody (A15505-1). NPTXR was detected in a paraffin-embedded section of mouse brain tissue. Heat me
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000221890-NPTXR/tissue/cerebral+cortex#imid_479
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000221890-NPTXR/tissue/cerebral+cortex#imid_20068917
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000221890-NPTXR/tissue/colon#imid_20068835
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000221890-NPTXR/tissue/liver#imid_20068844
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA001079; green = NPTXR, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA001079; green = NPTXR, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NPTXR IHC & IF
🧪Run cerebral cortex first and look for NPTXR staining in neuronal cells (HPA: Medium in cerebral cortex neuronal cells). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the cortex slide, use morphologically identified cells with background-only staining as internal comparators without assuming all non-neuronal cells are negative.
Positive control tissue: Cerebral cortex (Neuronal cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NPTXR in RT-4, SiHa, U2OS, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality, and NPTXR-knockout tissue or cells as a biological negative (selected tissue-IHC caption: rabbit primary antibody). For chromogenic IHC, quench endogenous peroxidase and check the cerebral cortex control slide for background staining (selected tissue-IHC caption: HRP/DAB detection); for IF/ICC, assess tissue autofluorescence with a no-primary control (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A15505-1 paraffin-section caption does not state the fixative (selected tissue-IHC caption: fixative not stated). The caption documents heat retrieval in EDTA at pH 8.0 and a primary concentration of 2 μg/ml, but does not establish that retrieval is required under other conditions (selected tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; for cerebral cortex IF, assess autofluorescence as a potential background artefact (HPA: Medium in cerebral cortex neuronal cells; standard IF practice).
HPA tissue IHC evidence for NPTXR
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | Medium | 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 NPTXR IHC Tips
Troubleshoot NPTXR staining in paraffin sections by checking retrieval, tissue morphology, cellular location, and controls before comparing chromogenic signal.
How should I adjust retrieval when NPTXR staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A15505-1). This condition accompanied NPTXR staining in human glioma tissue with 2 μg/ml primary antibody overnight at 4°C (datasheet A15505-1). If signal is weak, compare heating durations on adjacent sections while holding antibody incubation and DAB development constant (standard IHC practice). Check morphology as well as signal, since excessive retrieval can damage sections and increase background (standard IHC practice). Include cerebral cortex as a neuronal comparator and a section without primary antibody to assess detection background (HPA: medium staining in cortical neuronal cells; standard IHC practice).
Can fixation explain weak NPTXR staining in paraffin sections?
NPTXR-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not report its fixative (datasheet A15505-1). Record fixation conditions for each specimen and compare sections processed together before attributing a signal difference to NPTXR abundance (standard IHC practice). Inspect morphology for processing damage, then repeat the same EDTA pH 8.0 retrieval and antibody incubation across the comparison (datasheet A15505-1; standard IHC practice). A cerebral cortex section can help reveal a failed staining run because neuronal cells showed medium staining there (HPA: cerebral cortex neuronal cells, Medium). Fixation effects require a controlled comparison using this antibody (standard IHC practice).
Which compartments should count as plausible NPTXR staining?
Assess membrane-associated and cytoplasmic staining together: NPTXR is a membrane protein with extracellular residues 24–500, while cell imaging reports plasma membrane and cytosol (UniProt O95502 topology; HPA: approved subcellular locations). Tissue IHC describes cytoplasmic expression in a subset of cerebral cortex neuronal cells (HPA: tissue IHC profile). Nucleoplasmic signal has also been reported in cell imaging, so nuclear color alone warrants comparison with tissue controls before interpretation (HPA: additional nucleoplasmic location; standard IHC practice). Review individual cells and their morphology instead of scoring every stained area as neuronal (standard IHC practice). Compare staining with an adjacent negative-primary section and the expected cortical neuronal pattern (HPA: cerebral cortex; standard IHC practice).
How does epitope placement affect NPTXR IHC interpretation?
Check the antibody's documented immunogen or epitope before interpreting compartment differences; no antibody epitope is supplied here (datasheet A15505-1: epitope not stated). NPTXR has a short cytoplasmic segment at residues 1–2 and an extracellular region at 24–500 (UniProt O95502 topology). Its pentraxin domain spans 292–494, with annotated glycosylation sites at 42, 216, and 463 (UniProt O95502 domains and glycosylation). Those features locate possible epitope environments but do not establish a fixation or retrieval effect for this antibody (UniProt O95502; datasheet A15505-1: epitope not stated). No isoforms are listed, so an isoform-specific staining explanation lacks support in this record (UniProt O95502: isoforms 0).
How should I troubleshoot NPTXR in a multiplex IF experiment?
Treat IF/ICC as a separate assay: the supplied product image documents chromogenic staining of a paraffin section, while cell imaging reports membrane and cytosolic NPTXR (datasheet A15505-1; HPA: approved subcellular locations). Multiplex NPTXR with a neuronal marker when examining cerebral cortex, where a subset of neuronal cells shows staining (HPA: tissue IHC profile; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence, and use separate single-label controls to check channel bleed-through (standard IF practice). Determine the antibody epitope before selecting permeabilisation: an intracellular epitope requires access through the membrane, while an extracellular epitope may be assessed without it (UniProt O95502 topology; standard IF practice). The epitope and IF-specific fixation conditions are not supplied for this antibody (datasheet A15505-1).
What should I check when NPTXR DAB staining is diffuse?
First compare the stained section with a primary-omission control to identify color arising from the detection system (standard IHC practice). Block endogenous peroxidase before HRP detection, and check whether DAB development time produces color in control sections (standard chromogenic IHC practice). The selected example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and an HRP/DAB readout (datasheet A15505-1). If background persists, compare a lower primary concentration and shorter DAB development on adjacent sections while preserving the same retrieval condition (standard IHC practice; datasheet A15505-1: EDTA pH 8.0). Judge retained signal against cellular morphology and the cortical neuronal pattern (HPA: tissue IHC profile; standard IHC practice).
How should I quantify NPTXR staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring: cortical tissue IHC reports cytoplasmic staining in a subset of neuronal cells (HPA: tissue IHC profile). For chromogenic sections, record the percentage of positive target cells and staining intensity, then calculate an H-score if the intensity scale is defined consistently (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue or a specified anatomical region (standard IHC practice). Keep retrieval, antibody incubation, imaging, and DAB development consistent across samples; the documented retrieval is EDTA pH 8.0 (datasheet A15505-1; standard IHC practice). Normalize counts to the eligible cell population or analyzed tissue area, and exclude damaged regions using predefined rules (standard IHC practice).
How can I distinguish true NPTXR staining from artefact?
Look for a reproducible cellular pattern compatible with membrane or cytosolic NPTXR, including the reported cytoplasmic staining of some cortical neuronal cells (UniProt O95502: membrane; HPA: subcellular and tissue IHC). Check staining in the wrong cell population or an unexpected compartment against morphology and an independently processed comparator (standard IHC practice). Edge effects and necrotic areas can create misleading DAB deposits, so inspect them separately from intact tissue (standard IHC practice). A primary-omission control and endogenous-peroxidase block help identify detection-related color (standard chromogenic IHC practice). The glioma image establishes staining under its stated conditions; it does not establish that every glioma cell is NPTXR-positive (datasheet A15505-1).
Boster reagents
Best NPTXR / Neuronal pentraxin receptor IHC Antibodies
A15505-1 has real IHC data from paraffin sections of human glioma, mouse brain and rat brain (catalog image captions). No IF/ICC data are supplied (catalog applications and images).
Real IHC data
Anti-NPTXR Antibody ®
Cat # A15505-1
A15505-1 will render with its IHC figure from a paraffin section of human glioma (catalog image caption). Its additional captions show mouse and rat brain paraffin sections; the catalog lists human, mouse and rat reactivity (catalog image captions; catalog reactivity).
Which to pick: Choose A15505-1 for tissue IHC: its captions document paraffin sections processed with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog image captions). The same SKU is the cross-species choice for human, mouse and rat IHC (catalog reactivity; catalog image captions); the fixative is unreported (catalog image captions). No IF/ICC SKU can be recommended from this payload because IF/ICC is absent from the application list and no IF images are supplied (catalog applications and images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95502 (NPTXR_HUMAN, Neuronal pentraxin receptor).
- Human Protein Atlas. NPTXR tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NPTXR subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. NPTXR antibody validation summary (1 antibodies).
- Therapeutic monoclonal antibody targeting of neuronal pentraxin receptor to control metastasis in gastric cancer. Molecular cancer 2020 — PMC7448342.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NPTXR (Neuronal pentraxin receptor). Expected localisation Cytoplasmic in a subset of cortical neurons (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A15505-1). Validated IHC antibody: A15505-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioma tissue; fixative not specified (datasheet A15505-1); 2. Fixation: Image fixative and duration unreported (datasheet A15505-1); 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 A15505-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A15505-1); 8. Primary antibody: Rabbit anti-NPTXR, 2-5 μg/ml (datasheet A15505-1); 9. Primary incubation: Overnight at 4 °C (datasheet A15505-1). Watch for: Fixation: Keep paraffin-section fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Only a subset of cortical neurons stains (HPA tissue IHC); Isoform / epitope: No isoforms; extracellular 24–500 vs cytoplasmic 1–2 matters (UniProt); Regulation: Brain-enriched RNA expression (HPA RNA). Evidence: UniProt O95502, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A15505-1, PMC7448342. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NPTXR/immunohistochemistry-ihc-guide.