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NVL / Nuclear valosin-containing protein-like · IHC design guide
Design Immunohistochemistry for NVL
Plan paraffin-section NVL IHC using the nucleolar and cytoplasmic tissue pattern (HPA tissue IHC). Kidney distal tubules show high staining (HPA tissue IHC); check epitope coverage across the 5 annotated isoforms (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NVL 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 | IHC: nucleolar/cytoplasmic (HPA tissue IHC); protein: predominantly nucleolar (UniProt) | |
| Staining pattern | Nucleolar/cytoplasmic in several tissues; high in kidney distal tubules (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05446-3) | |
| Positive control | Kidney+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 compared paraffin sections. (selected-SKU IHC image A05446-3) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Specific expression regulation not reported (UniProt) | |
| Isoform / epitope | 5 isoforms; isoform-specific epitope coverage unknown (UniProt) |
Section 1
Recommended NVL IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A05446-3) is accompanied by one published NVL IHC protocol in human prostate adenocarcinoma sections (PMC12220030).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05446-3) |
| Fixation | Image fixative and duration unreported (datasheet A05446-3); 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 A05446-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05446-3) |
| Primary antibody | Rabbit anti-NVL, 2-5 μg/ml (datasheet A05446-3) |
| Primary incubation | Overnight at 4 °C (datasheet A05446-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05446-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NVL-positive staining in distal tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Nucleolar and cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A05446-3). The published method lists citrate pH 6.0 or EDTA pH 9.0 without specifying which was used for NVL (PMC12220030).
Section 2
What Is the Expected NVL Staining Pattern?
NVL is predominantly nucleolar, with nucleoplasmic localization also reported (UniProt O15381); it has no transmembrane segment (UniProt O15381 topology). In paraffin-section IHC, expect nucleolar staining in some cells, with cytoplasmic staining also reported across several tissues (HPA tissue IHC). Distal tubular cells in kidney show the strongest listed staining; several epithelial cell types show medium staining (HPA tissue IHC). The tissue IHC assessment is Supported, with medium consistency between staining and RNA data (HPA tissue IHC).
| Distinct nucleolar staining in kidney distal tubules, with or without cytoplasmic staining. | This fits the strongest listed tissue result, High staining in distal tubules (HPA tissue IHC), and the predominant nucleolar localization (UniProt O15381; HPA subcellular ICC-IF). Assess the nuclear compartment and cell identity together; staining intensity alone does not establish specificity (general IHC interpretation). |
| Strong, uniform staining confined to membranes or extracellular material, without recognizable nucleolar signal. | Treat this as a suspect compartment pattern: NVL is predominantly nucleolar and has no transmembrane segment (UniProt O15381 topology; HPA subcellular ICC-IF). Check the control slide and detection system before interpreting it as NVL; the supplied sources do not establish a membrane or extracellular NVL pattern. |
| Strong staining in adipocytes or cardiomyocytes while kidney distal tubules stain as expected. | Consider cross-reactivity or endogenous detection activity, then review whether staining truly belongs to those cells (general IHC interpretation). HPA lists NVL as Not detected in adipocytes and cardiomyocytes (HPA tissue IHC). A single unexpected positive cell population cannot by itself identify the cause. |
| Diffuse color across many compartments, including areas without a discrete nucleolar pattern. | This is difficult to score as specific NVL staining. HPA describes nucleolar and cytoplasmic expression in several tissues (HPA tissue IHC), so cytoplasmic color alone is insufficient evidence of artefact. Compare the distribution with a detection control and a kidney distal-tubule section before changing the interpretation (general IHC practice). |
| No detectable staining in kidney distal tubules. | A technical false negative is possible because those cells are listed as High (HPA tissue IHC). Check section integrity, primary-antibody conditions, retrieval and detection controls as general IHC practice. Absence of signal does not establish that NVL is biologically absent from the sampled kidney. |
💡Expected NVL appearanceCall a positive result when kidney distal tubular cells show High staining with a discernible nucleolar component (HPA tissue IHC; UniProt O15381); strong membrane-only, extracellular or broadly diffuse color without that component warrants artefact review (general IHC interpretation).
| Compartment and cell identity | Predominant nucleolar localization provides the main compartment check (UniProt O15381; HPA subcellular ICC-IF). HPA tissue IHC also reports cytoplasmic expression in several tissues, so cytoplasmic staining should be judged alongside nucleolar signal, cell identity and controls, not rejected automatically. |
| Tissue choice and source disagreement | Kidney distal tubules are a useful positive reference because HPA scores them High; breast and gallbladder glandular cells are among the listed Medium examples (HPA tissue IHC). UniProt reports high expression in heart, yet HPA scores cardiomyocytes Not detected (UniProt O15381 tissue specificity; HPA tissue IHC). Do not turn either observation into a universal tissue prediction. |
| Antibody validation | HPA reports IHC Supported for HPA028207, HPA028224 and HPA028654 (HPA antibodies). That status supports use of their recorded IHC patterns, but the tissue profile has only medium consistency with RNA data (HPA tissue IHC). Antibody-specific results should be interpreted within their own validation and staining context. |
| Isoforms and processing | UniProt lists 5 NVL isoforms and a full-length chain of residues 1–856, without a signal peptide or propeptide (UniProt O15381). These facts do not identify the epitope recognized by the chosen IHC antibody or predict which isoforms it detects; make no isoform-specific staining claim without antibody-specific evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney distal tubules are blank. | Possible section, retrieval, antibody or detection failure; the cause is not established by the blank result (general IHC practice). | Verify tissue morphology and detection controls, then optimize retrieval and primary-antibody conditions using the selected antibody’s IHC instructions (general IHC practice). Kidney distal tubules are the listed High reference (HPA tissue IHC). |
| Color is widespread and obscures nuclei. | Background from the detection workflow or nonspecific binding is possible (general IHC practice). HPA also reports cytoplasmic NVL staining in several tissues (HPA tissue IHC). | Run an appropriate primary-omission or detection control, review blocking and washes, and reassess whether any discrete nucleolar pattern remains (general IHC practice). Do not equate all cytoplasmic staining with background. |
| The dominant signal appears membranous. | The compartment conflicts with predominant nucleolar localization and the absence of a transmembrane segment (UniProt O15381; HPA subcellular ICC-IF). | Confirm the stained structure at higher magnification and compare with a positive tissue and detection control (general IHC practice). Avoid scoring membrane-only signal as NVL on this evidence. |
| Adipocytes or cardiomyocytes stain strongly. | Cross-reactivity, endogenous detection activity or cell misidentification is possible (general IHC interpretation); HPA scores those cells Not detected (HPA tissue IHC). | Check cell identity and control staining, then compare the result with kidney distal tubules on the same run (general IHC practice; HPA tissue IHC). Treat the unexpected result as unresolved until verified. |
| A tissue expected from expression summaries gives weak or no staining. | Tissue-wide expression and cell-specific IHC can disagree: UniProt lists high heart expression, while HPA reports cardiomyocytes Not detected (UniProt O15381 tissue specificity; HPA tissue IHC). | Record the exact cell population and assay result. Use a listed HPA-positive cell population as the IHC reference, and avoid assigning a target-specific fixation or retrieval explanation without evidence. |
| Can the IHC pattern be applied directly to IF/ICC? | The assays show related localization evidence, but HPA tissue IHC reports nucleolar and cytoplasmic expression, while HPA ICC-IF reports mainly nucleolar localization (HPA tissue IHC; HPA subcellular ICC-IF). | Use the IF/ICC guide for that assay. Here, regard a nucleolar signal as the shared localization check; do not treat an IHC chromogenic intensity or tissue score as an IF/ICC acceptance threshold (general assay interpretation). |
Experiment evidence
IHC & IF images showing NVL staining
A05446-3 · IHCIHC analysis of NVL using anti-NVL antibody (A05446-3). NVL was detected in a paraffin-embedded section of human liver cancer tissue. Heat m
A05446-3 · IHCIHC analysis of NVL using anti-NVL antibody (A05446-3). NVL was detected in a paraffin-embedded section of human colorectal adenocarcinoma t
A05446-3 · IFIF analysis of NVL using anti-NVL antibody (A05446-3) and anti-Beta Tubulin antibody (M01857-3). NVL was detected in immunocytochemical sect
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000143748-NVL/tissue/cerebral+cortex#imid_18748648
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000143748-NVL/tissue/colon#imid_18748558
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000143748-NVL/tissue/liver#imid_18748575
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000143748-NVL/tissue/kidney#imid_18748568
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA028219; green = NVL, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA028219; green = NVL, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NVL IHC & IF
🧪Run kidney first and look for staining in distal tubule cells (HPA: High in kidney distal tubules); use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes). On the kidney slide, cells without specific nucleolar staining can serve as internal negative comparators, but the supplied HPA row does not identify a particular negative kidney cell type (HPA: High in distal tubules; HPA subcellular: Nucleoli enhanced).
Positive control tissue: Kidney (Distal tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NVL in HEK293, Rh30, U2OS, MCF-7, with annotated localisation: Nucleoli (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody); use NVL knockout material or a validated peptide block as a biological specificity control. Quench endogenous peroxidase and check background in kidney before interpreting DAB signal (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the exact selected-SKU caption does not state a fixative; its paraffin section therefore does not establish fixation (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is documented for the selected SKU in human liver cancer tissue, but retrieval dependence in kidney is unreported (selected-SKU caption: EDTA pH 8.0). Frozen sections or IF cannot be judged easier from the supplied evidence; for IF/ICC, nucleolar localisation offers a staining pattern to assess, while kidney IHC requires attention to endogenous peroxidase background (HPA subcellular: Nucleoli enhanced; standard IHC practice). The selected A05446-3 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A05446-3).
HPA tissue IHC evidence for NVL
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Kidney | Distal tubules | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Medium | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NVL IHC Tips
Troubleshoot chromogenic NVL IHC in paraffin sections by checking retrieval, compartment-specific staining, controls and scoring before interpreting differences between samples.
How should I retrieve NVL in paraffin sections when nucleolar staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A05446-3). The selected tissue image used this retrieval before an overnight incubation at 4°C with 2 μg/ml primary antibody, providing a starting condition for this SKU (caption A05446-3). If staining remains weak, compare retrieval times on adjacent sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Score nucleolar signal separately from diffuse staining because NVL is predominantly nucleolar, although nucleoplasmic localization is also reported (UniProt O15381; HPA subcellular). Include a no-primary control to expose retrieval-associated background (standard IHC practice).
Can I infer which fixative best preserves the NVL epitope?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A05446-3). Do not assign its EDTA retrieval result to a particular fixation method, or infer fixation tolerance from NVL localization or modifications (caption A05446-3; UniProt O15381). For a fixation comparison, process matched specimens with recorded fixatives and fixation times, then stain adjacent sections using EDTA pH 8.0 retrieval and the same detection run (datasheet A05446-3; standard IHC practice). Compare nucleolar signal, morphology and no-primary background before choosing a condition (UniProt O15381; standard IHC practice). Record the chosen condition for subsequent batches (standard IHC practice).
What staining pattern should I expect, and how should I assess cytoplasmic signal?
Expect the clearest NVL signal in nucleoli, with possible nucleoplasmic staining (UniProt O15381; HPA subcellular). Tissue IHC also reports nucleolar and cytoplasmic expression in several tissues, with supported reliability and medium agreement between staining and RNA expression (HPA tissue IHC). Assess each compartment separately against an identifiable nuclear counterstain, and compare cytoplasmic signal with no-primary and adjacent-section controls (standard IHC practice). In kidney, distal tubules are a useful positive reference because their staining is reported as high (HPA tissue IHC). Treat isolated cytoplasmic staining without corresponding controls or reproducible cell-specific pattern as unresolved, rather than assigning it automatically to NVL (HPA tissue IHC; standard IHC practice).
Could isoforms or modifications explain inconsistent NVL staining across sections?
NVL has 5 annotated isoforms, while the supplied catalog evidence does not identify the antibody epitope or establish isoform coverage (UniProt O15381; caption A05446-3). Annotated modifications include acetylation at residue 70 and phosphorylation at residues 134 and 138; their effects on this antibody's staining are unknown (UniProt O15381). Check the antibody's epitope information before interpreting a negative section as absence of every isoform (standard IHC practice). Compare matched sections with the same EDTA pH 8.0 retrieval and detection settings to isolate processing variation (datasheet A05446-3; standard IHC practice). Report the antibody and observed compartment alongside any isoform-level hypothesis (standard IHC practice).
How can IF help verify the cellular source of NVL IHC staining?
On a matched section, multiplex NVL with a validated marker for the suspected cell type, such as distal tubular cells when assessing kidney, and compare the same cells across channels (HPA tissue IHC; standard IF practice). Choose a red or far-red fluorophore after checking the tissue's autofluorescence in each channel (standard IF practice). Use a permeabilization condition that permits antibody access to nuclear compartments, because NVL is mainly nucleolar and has no annotated transmembrane segment (UniProt O15381; standard IF practice). Include single-label and no-primary controls to assess bleed-through and background (standard IF practice). Treat IF as a localization cross-check: the selected catalog caption documents paraffin-section chromogenic IHC, not an IF protocol (caption A05446-3).
What should I check when DAB obscures nucleolar NVL staining?
The selected assay used 10% goat serum blocking, a peroxidase-conjugated secondary antibody and DAB development (caption A05446-3). If diffuse brown signal obscures nucleoli, compare no-primary and secondary-only controls, inspect section edges, and shorten development or titrate the primary on adjacent sections (standard IHC practice). Include an endogenous peroxidase block and verify its effectiveness with an appropriate control; this is a general chromogenic IHC step, not NVL-specific validation (standard IHC practice). Keep EDTA pH 8.0 retrieval constant while adjusting detection so changes remain interpretable (datasheet A05446-3; standard IHC practice). Evaluate background within the same cell compartment used for scoring (standard IHC practice).
How should I quantify NVL staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; nucleolar staining is the primary localization reference for NVL (UniProt O15381; HPA subcellular). For each specimen, record the percentage of positive eligible cells and intensity, then calculate an H-score using 0–3 intensity categories if that scale is prespecified (standard IHC practice). Alternatively, report positive cells per mm² when cell density is the biological endpoint (standard IHC practice). Normalize counts to the number of eligible, intact cells or the evaluated viable tissue area, and use the same threshold across batches (standard IHC practice). Record cytoplasmic staining separately because tissue IHC reports it in several tissues (HPA tissue IHC).
When does an apparent NVL-positive result need further validation?
A convincing result shows reproducible staining in expected cellular compartments, especially nucleoli, alongside clean assay controls (UniProt O15381; HPA subcellular; standard IHC practice). Check the cell identity: kidney distal tubules have reported high staining, whereas adipocytes are reported as not detected in the cited tissue survey (HPA tissue IHC). Review staining confined to section edges or necrotic areas against adjacent viable tissue, since those patterns can reflect preparation artefacts (standard IHC practice). Test brown signal in no-primary sections to identify detection background, including endogenous enzyme activity (standard IHC practice). Interpret unexpected cytoplasmic signal cautiously because tissue IHC reports it, but its agreement with RNA data is only medium (HPA tissue IHC).
Boster reagents
Best NVL / Nuclear valosin-containing protein-like IHC Antibodies
The IHC-validated anti-NVL antibody has paraffin-section data from human liver cancer tissue and IF/ICC data from U2OS cells (catalog image captions); its listed reactivity is human and rat (catalog).
Real IHC data
Anti-NVL Antibody ®
Cat # A05446-3
A05446-3 is listed for IHC and was shown on a paraffin-embedded human liver cancer section with EDTA retrieval and DAB detection (catalog IHC caption). A05446-3 is also listed for IF/ICC, with an IF image from U2OS cells (catalog IF caption).
Which to pick: Choose A05446-3 for human tissue IHC: its paraffin-section image used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC caption); the fixative was unreported (catalog IHC caption). For IF/ICC, the same rabbit antibody has a U2OS cell image at 5 μg/ml (catalog IF caption). For rat samples, A05446-3 lists rat reactivity for ICC/IF, while its listed paraffin-section IHC application and supplied IHC images are human (catalog dilution listing and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15381 (NVL_HUMAN, Nuclear valosin-containing protein-like).
- Human Protein Atlas. NVL tissue IHC expression (reliability: Supported).
- Human Protein Atlas. NVL subcellular location (ICC-IF): Mainly localized to the nucleoli..
- Human Protein Atlas. NVL antibody validation summary (4 antibodies).
- Construction and verification of a prognostic model for prostate cancer based on ribosome biogenesis-related genes. BMC medical genomics 2025 — PMC12717746.
- Construction of a prostate adenocarcinoma molecular classification: integrating spatial transcriptomics with retrospective cohort validation. Journal of translational medicine 2025 — PMC12220030.
- PubMed PMID:9286697 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NVL (Nuclear valosin-containing protein-like). Expected localisation IHC: nucleolar/cytoplasmic (HPA tissue IHC); protein: predominantly nucleolar (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05446-3). Validated IHC antibody: A05446-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Distal tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05446-3); 2. Fixation: Image fixative and duration unreported (datasheet A05446-3); 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 A05446-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05446-3); 8. Primary antibody: Rabbit anti-NVL, 2-5 μg/ml (datasheet A05446-3); 9. Primary incubation: Overnight at 4 °C (datasheet A05446-3). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A05446-3); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; isoform-specific epitope coverage unknown (UniProt); Regulation: Specific expression regulation not reported (UniProt). Evidence: UniProt O15381, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoli., Datasheet A05446-3, PMC12220030. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NVL/immunohistochemistry-ihc-guide.