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NAP1L5 / Nucleosome assembly protein 1-like 5 · IHC design guide
Design Immunohistochemistry for NAP1L5
Plan chromogenic IHC-P for NAP1L5 using its mainly nuclear tissue pattern, most abundant in CNS (HPA tissue IHC). Start the catalog antibody at 5 μg/mL (datasheet: A14492) and interpret staining in light of the reported antibody–RNA discordance (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NAP1L5 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 | Mainly nuclear in tissues (HPA tissue IHC) | |
| Staining pattern | Mainly nuclear across tissues; most abundant in CNS (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+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 A14492) | |
| Caveat | Low antibody–RNA concordance may complicate interpretation (HPA tissue IHC) | |
| Regulation | Brain-predominant expression (UniProt) | |
| Isoform / epitope | No isoforms or processing annotated; epitope map unresolved (UniProt) |
Section 1
Recommended NAP1L5 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published NAP1L5 staining protocol for carcinoma and tumor transplantation tissues (PMC9740361).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A14492); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-NAP1L5, 5 μg/mL (datasheet A14492) |
| 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 | NAP1L5-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Mainly nuclear expression in several tissues, most abundant CNS. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min for this nuclear antigen (UniProt Q96NT1; page retrieval rule). The article does not specify retrieval conditions (PMC9740361).
Section 2
What Is the Expected NAP1L5 Staining Pattern?
NAP1L5 is a nuclear protein without a transmembrane segment (UniProt Q96NT1: subcellular location and topology). In paraffin-section IHC, expect mainly nuclear staining, most abundant in CNS tissues; HPA reports medium staining in cortical and caudate neuronal cells and cerebellar molecular-layer cells (HPA: tissue IHC). HPA rates its tissue IHC profile Approved but reports low consistency between antibody staining and RNA expression, so interpret isolated positive cells cautiously (HPA: tissue IHC reliability).
| Distinct nuclear staining in cortical or caudate neuronal cells. | This matches HPA's medium staining in those cells and its mainly nuclear tissue profile (HPA: tissue IHC). Compare the signal with adjacent morphology and controls before scoring it as target-associated staining (general IHC practice). |
| Strong membrane staining, or cytoplasmic staining with no discernible nuclear component. | A dominant membrane pattern conflicts with the nuclear location and lack of transmembrane segment (UniProt Q96NT1: location and topology). Cytoplasmic signal alone is insufficient to establish the expected tissue IHC pattern; examine background and detection controls (HPA: tissue IHC; general IHC practice). |
| Prominent staining of adipocytes or liver cholangiocytes. | HPA reports these cells as not detected in tissue IHC (HPA: adipose tissue and liver IHC). Treat unexpected staining as a specificity or endogenous-detection question; inspect the negative control and tissue morphology before interpreting it as NAP1L5 (general IHC practice). |
| Diffuse color across nuclei, cytoplasm, and surrounding tissue. | A widespread haze does not resolve the mainly nuclear pattern reported by HPA (HPA: tissue IHC). Uneven blocking, excessive detection signal, or incomplete washing can cause nonspecific background; compare with a no-primary control (general IHC practice). |
| No staining in cortical or caudate neuronal cells. | HPA reports medium staining in these cells, so an absent result warrants a run check (HPA: tissue IHC). Confirm tissue preservation and morphology, then review the catalog antibody's IHC-P conditions and the run controls (general IHC practice). |
💡Expected NAP1L5 appearanceCall a convincing positive result mainly nuclear staining of cortical or caudate neuronal cells at about HPA's medium level; dominant membrane signal or diffuse tissue-wide color is discordant (HPA: tissue IHC; UniProt Q96NT1: topology).
| Cell type and tissue | Brain is the predominant expression site (UniProt Q96NT1: tissue specificity). HPA finds medium neuronal staining in cortex and caudate, but low staining in hippocampal neuronal cells; choose and score reference areas by cell type (HPA: tissue IHC). |
| Compartment | UniProt places NAP1L5 in the nucleus and reports no transmembrane segment (UniProt Q96NT1: location and topology). HPA describes tissue staining as mainly nuclear; membrane-dominant staining needs scrutiny (HPA: tissue IHC). |
| Evidence strength | HPA labels the tissue IHC profile Approved while noting low consistency with RNA expression (HPA: tissue IHC reliability). Its listed antibody HPA058227 is Approved for IHC, which does not resolve every unexpected cell-level result (HPA: antibody validation; HPA: tissue IHC reliability). |
| Protein processing | UniProt lists one 1–182 chain, no signal peptide or propeptide, and no annotated glycosylation sites or modified residues (UniProt Q96NT1: processing and modifications). These annotations give no basis to expect a secreted or membrane-associated staining pattern (UniProt Q96NT1: processing and topology). |
| IHC-P workflow variables | Antigen retrieval, blocking, primary-antibody concentration, washing, and chromogenic detection can affect contrast in routine paraffin-section IHC (general IHC practice). Target-specific fixation sensitivity and an optimal retrieval condition are unreported in the supplied UniProt and HPA records. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclear signal is weak in an HPA-positive reference area. | Cortical and caudate neuronal cells have medium HPA staining, so weak signal may reflect the selected cells or the run conditions (HPA: tissue IHC; general IHC practice). | Check neuronal morphology and run controls, then review the catalog antibody's IHC-P retrieval and detection instructions before adjusting routine conditions (general IHC practice). |
| Most cells show brown haze. | Widespread color obscures HPA's mainly nuclear tissue pattern and may reflect nonspecific detection or inadequate washing (HPA: tissue IHC; general IHC practice). | Compare a no-primary control, inspect blocking and washes, and reduce detection intensity if the control also develops color (general IHC practice). |
| Signal appears mainly at cell membranes. | Membrane dominance conflicts with NAP1L5's nuclear location and absence of a transmembrane segment (UniProt Q96NT1: location and topology). | Check morphology and control sections; do not score membrane-only color as the expected NAP1L5 pattern (HPA: tissue IHC; general IHC practice). |
| A nominally negative cell population stains strongly. | HPA reports adipocytes and liver cholangiocytes as not detected; cross-reactivity or endogenous detection activity may explain unexpected color (HPA: tissue IHC; general IHC practice). | Check the no-primary control and localize color to specific cell types before assigning target expression (general IHC practice). |
| Cortex is positive but hippocampal neuronal cells are faint. | This difference can match HPA's medium cortical and low hippocampal neuronal staining (HPA: tissue IHC). | Score the regions separately and compare like cell types; faint hippocampal signal alone does not establish a failed run (HPA: tissue IHC; general IHC practice). |
| What should a separate IF/ICC experiment show? | HPA places NAP1L5 mainly in the nucleoplasm, with an additional cytosolic location in ICC-IF (HPA: subcellular ICC-IF). | Use the separate IF/ICC guide for that application; compare nuclear and cytosolic fluorescence with appropriate controls rather than applying the tissue IHC pattern as an exclusive ICC-IF rule (HPA: subcellular ICC-IF; general IF practice). |
Experiment evidence
IHC & IF images showing NAP1L5 staining
A14492 · IHCImmunohistochemistry of NAP1L5 in mouse brain tissue with NAP1L5 antibody at 5 μg/mL.
A14492 · IFImmunofluorescence of NAP1L5 in mouse brain tissue with NAP1L5 antibody at 20 μg/mL.
A14492-1 · IFIF analysis of NAP1L5 using anti-NAP1L5 antibody (A14492-1) and anti-Beta Tubulin antibody (M01857-3). NAP1L5 was detected in immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000177432-NAP1L5/tissue/cerebral+cortex#imid_19937285
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000177432-NAP1L5/tissue/colon#imid_19937211
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000177432-NAP1L5/tissue/liver#imid_19937212
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000177432-NAP1L5/tissue/kidney#imid_19937221
![HPA ICC-IF BJ [Human fibroblast] — NAP1L5 (HPA058227)](https://images.proteinatlas.org/58227/1599_B11_3_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA058227; green = NAP1L5, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — NAP1L5 (HPA058227)](https://images.proteinatlas.org/58227/1599_B11_5_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA058227; green = NAP1L5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NAP1L5 IHC & IF
🧪Run cerebral cortex first and assess neuronal cells for NAP1L5 staining (HPA: Medium in cerebral cortex neuronal cells); use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes). On the positive slide, cells without specific signal should show counterstain without nuclear chromogen; do not assume an unlisted cell type is a validated negative control.
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NAP1L5 in BJ [Human fibroblast], SH-SY5Y, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and knockout tissue as a biological negative. For chromogenic IHC, quench endogenous peroxidase and distinguish any native brain pigment from reaction product (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; optimize retrieval empirically for paraffin sections. The selected A14492 mouse-brain IHC caption reports 5 μg/mL but leaves the fixative unreported (selected-SKU tissue-IHC caption). The supplied evidence does not establish that frozen sections or tissue IF are easier; brain autofluorescence can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for NAP1L5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | Medium | Protein (IHC) | HPA → |
| 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NAP1L5 IHC Tips
Troubleshoot NAP1L5 staining in paraffin section IHC using nuclear localisation and brain enriched expression as reference points (UniProt Q96NT1; HPA tissue IHC).
How should I adjust retrieval when nuclear NAP1L5 staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval setting: nuclear antigen). Compare a retrieved section with an otherwise matched section processed without heat, keeping antibody and detection conditions constant (standard IHC practice). Examine nuclear signal in brain alongside tissue integrity: NAP1L5 is reported in the nucleus, and brain is enriched for its RNA (UniProt Q96NT1; HPA tissue IHC). If signal remains weak, test a shorter or longer heating time on adjacent sections and assess morphology before changing other variables (standard IHC practice). Do not attribute weak staining to retrieval alone without a working positive control (standard IHC practice).
Could fixation explain absent NAP1L5 staining in paraffin sections?
The supplied mouse brain IHC caption does not state a fixative, so target specific fixation sensitivity is unknown (catalog antibody A14492 caption). Record fixation conditions for each specimen and compare similarly processed sections before assigning a biological difference to staining intensity (standard IHC practice). Run a matched positive brain control through the same fixation, processing, retrieval and detection steps; the caption documents mouse brain staining at 5 µg/mL (catalog antibody A14492 caption). If both control and test sections are weak, assess processing and retrieval systematically using the page setting of Tris-EDTA pH 9.0 for 20 min (page retrieval setting). A positive control cannot establish that all fixation conditions preserve the epitope equally (standard IHC practice).
Should cytoplasmic staining count as NAP1L5 positivity?
Prioritise nuclear staining when reading chromogenic sections: UniProt assigns NAP1L5 to the nucleus, and HPA describes mainly nuclear tissue staining (UniProt Q96NT1; HPA tissue IHC). HPA imaging also reports an approved nucleoplasmic location with an additional cytosolic location, so cytoplasmic signal is possible but requires careful control comparison (HPA subcellular). Score nuclear and cytoplasmic compartments separately rather than combining them into a single positive call (standard IHC practice). Compare the candidate pattern with omission of primary antibody and with a consistently processed brain control, where expression is enriched (standard IHC practice; HPA tissue IHC). Diffuse staining over tissue edges or damaged areas should be reviewed as a possible preparation artefact (standard IHC practice).
How can I assess an unexpected staining pattern without epitope mapping?
The record lists a 182 amino acid chain and 0 annotated isoforms, but supplies no antibody epitope, so the stained region cannot be assigned from these data (UniProt Q96NT1). It also lists no signal peptide, transmembrane segment or annotated glycosylation sites; these features do not establish how a particular antibody behaves in paraffin sections (UniProt Q96NT1). Compare adjacent sections across retrieval conditions while keeping the detection system fixed, then inspect whether nuclear signal changes together with background (standard IHC practice). If an epitope claim matters to the experiment, obtain antibody specific mapping or independent validation before interpreting an unusual compartment (standard IHC practice). Document the catalog antibody identity, retrieval setting and scored compartment for reproducibility (standard IHC practice).
How should I investigate the NAP1L5 pattern with multiplex IF?
Use IF as a secondary check by pairing NAP1L5 with a neuronal marker in brain sections, since HPA reports neuronal staining in caudate and cerebral cortex (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and favour a far red channel when brain autofluorescence obscures shorter wavelengths; inspect unstained tissue in each channel first (standard IF practice). Apply permeabilisation appropriate for access to intracellular nuclear and cytosolic epitopes, then check whether it preserves morphology (UniProt Q96NT1; HPA subcellular; standard IF practice). Include single stain controls to assess channel bleed through and a no primary control to assess background (standard IF practice). HPA reports nucleoplasmic and additional cytosolic localisation, which can guide comparison without requiring perfect overlap with the neuronal marker (HPA subcellular).
What should I check when DAB appears broadly positive?
First inspect a no primary control and an adjacent section stained with the catalog antibody under matched detection conditions (standard IHC practice). For chromogenic detection, block endogenous peroxidase before applying a peroxidase based system and develop DAB consistently across comparison sections (standard IHC practice). Check whether colour concentrates at tissue edges, folds or damaged regions, and distinguish these deposits from staining within intact nuclei (standard IHC practice; UniProt Q96NT1). If background persists, review blocking, washes, antibody concentration and development time one variable at a time; the supplied mouse brain caption reports 5 µg/mL for antibody A14492 (standard IHC practice; catalog antibody A14492 caption). Treat widespread signal in a no primary control as a detection problem until resolved (standard IHC practice).
How should I quantify NAP1L5 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and eligible cell population before scoring, then keep those rules constant across sections (standard IHC practice). Report the percentage of nuclei with convincing chromogenic signal and, when intensity grading is reliable, an H-score using 0–3 intensity categories (standard IHC practice). Normalise positive counts to the number of eligible nuclei or sampled area, such as cells per mm², and report the sampled area (standard IHC practice). Score cytoplasmic staining separately because HPA reports mainly nuclear tissue expression while cell imaging also shows an additional cytosolic location (HPA tissue IHC; HPA subcellular). Compare specimens processed in the same staining run and retain a brain control, given brain enriched expression (standard IHC practice; UniProt Q96NT1).
Which findings make an apparent NAP1L5 positive result doubtful?
Give greatest weight to staining in intact nuclei within a plausible cell population: NAP1L5 is nuclear, and HPA reports mainly nuclear tissue staining with prominent CNS expression (UniProt Q96NT1; HPA tissue IHC). Review isolated cytoplasmic staining, unexpected cell types, edge accentuation and necrotic areas against matched controls before calling them positive (standard IHC practice; HPA subcellular). Signal retained in a no primary control can indicate endogenous enzyme activity or another detection artefact in chromogenic IHC (standard IHC practice). HPA lists its tissue antibody as Approved but notes low consistency between staining and RNA expression, so staining alone warrants cautious biological interpretation (HPA tissue IHC). Record the compartment, cell type and control behaviour with each conclusion (standard IHC practice).
Boster reagents
Best NAP1L5 / Nucleosome assembly protein 1-like 5 IHC Antibodies
Two anti-NAP1L5 antibodies have mouse brain tissue IHC/IF images and a human HeLa cell IF/ICC image (catalog image captions). Both list human, mouse, and rat reactivity (catalog reactivity).
Real IHC data
Anti-NAP1L5 Antibody
Cat # A14492
Real IF data
Anti-NAP1L5 Antibody ®
Cat # A14492-1
A14492 lists IHC-P and IF, with mouse brain tissue images at 5 μg/mL for IHC and 20 μg/mL for IF (catalog applications and A14492 image captions). A14492-1 lists IF/ICC and has a HeLa cell IF image at 5 μg/mL (catalog applications and A14492-1 IF caption).
Which to pick: For paraffin-section tissue IHC, choose A14492: IHC-P is listed, and its own IHC caption shows mouse brain tissue at 5 μg/mL; the fixative is unreported (catalog applications and A14492 IHC caption). For cell IF/ICC, choose A14492-1, whose IF caption shows HeLa cells at 5 μg/mL (catalog applications and A14492-1 IF caption). Both list human, mouse, and rat reactivity; the pictured IHC evidence is mouse tissue for A14492, while the pictured cell IF evidence is human HeLa cells for A14492-1 (catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96NT1 (NP1L5_HUMAN, Nucleosome assembly protein 1-like 5).
- Human Protein Atlas. NAP1L5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NAP1L5 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. NAP1L5 antibody validation summary (1 antibodies).
- NAP1L5 targeting combined with MYH9 Inhibit HCC progression through PI3K/AKT/mTOR signaling pathway. Aging 2022 — PMC9740361.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:12383514 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NAP1L5 (Nucleosome assembly protein 1-like 5). Expected localisation Mainly nuclear in tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A14492 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A14492); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-NAP1L5, 5 μg/mL (datasheet A14492); 9. Primary incubation: Overnight at 4 °C (standard). 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 A14492); Caveat: Low antibody–RNA concordance may complicate interpretation (HPA tissue IHC); Isoform / epitope: No isoforms or processing annotated; epitope map unresolved (UniProt); Regulation: Brain-predominant expression (UniProt). Evidence: UniProt Q96NT1, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A14492, PMC9740361. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NAP1L5/immunohistochemistry-ihc-guide.