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NUP62 / Nuclear pore glycoprotein p62 · IHC design guide
Design Immunohistochemistry for NUP62
Plan NUP62 IHC in paraffin sections around the nuclear membrane staining seen in most cell types, with cytoplasmic staining in some (HPA tissue IHC). Use the catalog antibody’s IHC protocol and compare staining across sections with consistent fixation (datasheet A03950-2; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUP62 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear membrane in most cells; cytoplasm in some (HPA tissue IHC) | |
| Staining pattern | Most cells: nuclear membrane; some: cytoplasm (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03950-2) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Parathyroid glandular cells show low staining (HPA tissue IHC) | |
| Regulation | No specific regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; no transmembrane segment (UniProt) |
Section 1
Recommended NUP62 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA retrieval (datasheet: A03950-2). The published protocols below describe chromogenic NUP62 staining in rat and human tissue (PMC8169113; PMC11684083; PMC5595097).
| Sample | Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A03950-2) |
| Fixation | Image fixative and duration unreported (datasheet A03950-2); 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 A03950-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03950-2) |
| Primary antibody | Rabbit anti-NUP62, 2-5 μg/ml (datasheet A03950-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03950-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03950-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUP62-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Expression in the nuclear membranes in most cell types. Also cytoplasmic expression in several cases. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A03950-2); use each paper’s stated retrieval conditions when reproducing its protocol.
Section 2
What Is the Expected NUP62 Staining Pattern?
NUP62 is a nuclear pore protein with no transmembrane segment (UniProt P37198: localization and topology). In paraffin section IHC, expect staining along the nuclear membrane in most cell types; cytoplasmic staining can also occur (HPA: tissue IHC profile). HPA rates the tissue pattern Supported, citing high consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| A nuclear rim is visible in many cells, including strongly stained caudate neuronal cells, placental endothelial cells, or rectal glandular cells (HPA: tissue IHC, High). | This fits NUP62 at the nuclear pore complex (UniProt P37198: localization) and HPA's predominant nuclear membrane pattern (HPA: tissue IHC profile). Judge the rim against the counterstained nucleus; tissue labels alone do not establish the identity of every stained cell (general IHC practice). |
| Some cells show cytoplasmic staining in addition to a nuclear rim (HPA: tissue IHC profile). | Cytoplasmic signal is not automatically false: HPA reports it in several cases, and UniProt also places NUP62 at spindle poles and centrosomes (HPA: tissue IHC profile; UniProt P37198: localization). Assess whether the expected nuclear membrane pattern remains discernible before interpreting the extra signal. |
| Strong, uniform signal fills cells or extracellular areas while nuclear rims are indistinct. | Treat this as an uncertain pattern rather than evidence of NUP62 localization: the reported tissue pattern emphasizes nuclear membranes (HPA: tissue IHC profile). Diffuse chromogen, tissue pigment, or detection background can complicate compartment scoring (general IHC practice); inspect controls and morphology. |
| Staining appears mainly in an unexpected cell population, with little signal in the expected population. | Consider antibody cross-reactivity or endogenous detection activity (general IHC practice), but first check cell identification. HPA reports low staining in several sampled populations, not a list of negative tissues, and describes expression in most cell types (HPA: tissue IHC profile). A low-staining cell is therefore a weak exclusion control. |
| A known HPA High population has no discernible nuclear membrane staining. | The result needs a technical check before a biological absence call: HPA reports High staining in caudate neuronal, placental endothelial, and rectal glandular cells (HPA: tissue IHC). Check section quality, counterstain, antibody application, and detection with appropriate controls (general IHC practice). HPA staining levels do not establish fixation sensitivity. |
💡Expected NUP62 appearanceCall a result positive when an interpretable nuclear membrane rim appears in the expected cells, potentially strong in HPA High populations; isolated diffuse extracellular or uniform cytoplasmic chromogen without a discernible rim is suspect (HPA: tissue IHC profile and High populations; general IHC practice).
| Compartment and topology (UniProt P37198: localization and topology). | NUP62 occupies the nuclear pore complex and has no transmembrane segment (UniProt P37198). Score the nuclear boundary rather than assuming all nuclear or cytoplasmic color is equivalent; HPA's tissue pattern centers on the nuclear membrane (HPA: tissue IHC profile). |
| Cell and tissue context (HPA: tissue IHC). | Caudate neuronal cells, placental endothelial cells, and rectal glandular cells are High; appendix and breast glandular cells are Medium, among other listed Medium populations (HPA: tissue IHC). These are reported staining levels, not promised signal strengths for every section or run. |
| Cytoplasmic and mitotic context (HPA: tissue IHC; UniProt P37198: localization). | HPA reports cytoplasmic expression in several cases. UniProt describes association with mitotic spindle poles and centrosomes (HPA: tissue IHC profile; UniProt P37198: localization). Evaluate such staining with cell morphology and the nuclear rim pattern; neither source establishes that every cytoplasmic deposit is specific. |
| Evidence and antibody choice (HPA: antibody validation). | The listed antibodies HPA005435 and CAB020724 have Supported IHC status (HPA: antibody validation). Supported tissue reliability reflects agreement with RNA expression; it does not by itself prove every stained compartment in a new specimen is specific (HPA: tissue IHC reliability; general IHC practice). |
| Fixation evidence (HPA: tissue IHC; UniProt P37198). | No target-specific fixation sensitivity is supplied by these records. HPA staining levels and UniProt topology or modifications do not establish how fixation changes NUP62 detection (HPA: tissue IHC; UniProt P37198). Assess local processing with controls rather than assigning a target-specific fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control has no nuclear rim (HPA: tissue IHC pattern). | Antibody delivery, retrieval, detection, or section quality may have failed (general IHC practice); these sources do not identify a NUP62-specific fixation effect. | Repeat with a documented High population such as placental endothelial or rectal glandular cells (HPA: tissue IHC). Verify each IHC step and compare an appropriate run control (general IHC practice). |
| All cells or blank areas show diffuse chromogen. | Background from the detection workflow may obscure the nuclear boundary (general IHC practice). HPA's principal pattern is nuclear membrane staining (HPA: tissue IHC profile). | Inspect a no-primary control and tissue morphology; review blocking, washing, detection exposure, and counterstain using general IHC practice. Score only interpretable cellular staining. |
| Cytoplasm stains more strongly than the nuclear rim. | Some cytoplasmic NUP62 signal is plausible, while diffuse background is also possible (HPA: tissue IHC profile; general IHC practice). | Compare intact cells with the no-primary control and check whether a nuclear rim can still be resolved (HPA: tissue IHC profile; general IHC practice). Do not classify all cytoplasmic staining as artefact. |
| Only an HPA Low population is available as a comparison. | Low staining is not an absent-protein control; HPA lists low glandular staining in parathyroid and adrenal glands, among other Low populations (HPA: tissue IHC). | Add a documented High population to the same run where practical, and compare compartments within well-preserved cells (HPA: tissue IHC; general IHC practice). Avoid calling a Low population definitively negative. |
| A surprising cell population stains while the expected population does not. | Cell identification, cross-reactivity, or endogenous detection activity may account for the discrepancy (general IHC practice); HPA reports broad nuclear membrane expression (HPA: tissue IHC profile). | Recheck morphology and controls, then compare a listed High population and, if available, an independently validated antibody (HPA: tissue IHC and antibody validation; general IHC practice). |
| IF/ICC question: should fluorescence be read like the paraffin section IHC result? | HPA's ICC-IF summary places NUP62 mainly at the nuclear membrane and additionally in the nucleoplasm (HPA: subcellular ICC-IF); the tissue IHC profile also allows cytoplasmic staining in several cases (HPA: tissue IHC). | Use the nuclear membrane as the shared localization check, while interpreting nucleoplasmic fluorescence in the separate IF/ICC context (HPA: subcellular ICC-IF). Do not infer an IHC protocol choice from the ICC-IF images. |
Experiment evidence
IHC & IF images showing NUP62 staining
A03950-2 · IHCIHC analysis of NUP62 using anti-NUP62 antibody (A03950-2). NUP62 was detected in a paraffin-embedded section of human ovarian cancer tissue
A03950-2 · IHCIHC analysis of NUP62 using anti-NUP62 antibody (A03950-2). NUP62 was detected in a paraffin-embedded section of human ovarian cancer tissue
A03950-2 · IFIF analysis of NUP62 using anti-NUP62 antibody (A03950-2) and anti-Beta Tubulin antibody (M01857-3). NUP62 was detected in immunocytochemica
M03950 · IFIF analysis of NUP62 using anti-NUP62 antibody (M03950). NUP62 was detected in immunocytochemical section of Hela cell. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000213024-NUP62/tissue/cerebral+cortex#imid_5337864
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000213024-NUP62/tissue/colon#imid_5337806
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000213024-NUP62/tissue/liver#imid_5337791
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000213024-NUP62/tissue/pancreas#imid_5337758
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005435; green = NUP62, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005435; green = NUP62, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUP62 IHC & IF
🧪Run placenta first and score endothelial cells for NUP62 staining (HPA: High in placental endothelial cells). HPA lists no negative tissue and detects NUP62 in all 45 scored tissues, so no-primary and isotype controls provide the negative reference; no cell type on the positive slide is established as an internal antigen-negative control (HPA: negative rows empty; detected in all 45 scored tissues).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: None in HPA: NUP62 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUP62 in A-431, U-251MG, U2OS, hTERT-RPE1 (serum starved), with annotated localisation: Nuclear membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a section incubated with species- and clonality-matched rabbit IgG; where the cognate immunizing peptide is available, peptide-block the primary as a specificity control (selected-SKU caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and check blood-rich areas of placenta for background before interpreting DAB staining (standard IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but reports no fixative; a target-specific fixation window or fixation effect is unreported (selected-SKU tissue-IHC caption). IF/ICC can help assess the expected nuclear-envelope pattern, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA: supported nuclear-membrane localization; UniProt P37198: nuclear pore complex). In placenta, blood-associated endogenous peroxidase can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for NUP62
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Placenta | Endothelial cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: NUP62 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced NUP62 IHC Tips
Troubleshoot NUP62 staining in paraffin sections by checking retrieval, nuclear envelope localisation, controls and cell-specific scoring before interpreting signal.
What retrieval should I use when NUP62 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A03950-2). That condition was used for NUP62 staining in a paraffin section of human ovarian cancer tissue, although the caption does not state the fixative (caption A03950-2). Keep retrieval conditions identical across sections in a comparison, then check whether nuclear envelope staining improves without increased tissue damage (standard IHC practice; HPA: nuclear membrane is the main location). If signal remains weak, verify primary antibody concentration, detection reagents and a suitable positive control before changing retrieval conditions (standard IHC practice). The caption used 2 μg/ml primary antibody overnight at 4°C (caption A03950-2).
How should I troubleshoot suspected fixation-related loss of NUP62 staining?
The selected paraffin-section caption does not report a fixative, so target-specific NUP62 sensitivity to fixation is unknown (caption A03950-2). Record the fixative, fixation duration and tissue processing for each specimen, and compare sections processed under matched conditions when investigating weak staining (standard IHC practice). Use the reported EDTA retrieval at pH 8.0 as the starting condition and hold antibody concentration and detection steps constant while assessing processing differences (datasheet A03950-2; standard IHC practice). An intact nuclear counterstain alongside absent NUP62 signal does not establish a fixation effect; check a positive control and reagent performance first (standard IHC practice; HPA: nuclear membrane location).
Which staining pattern supports NUP62 localisation in chromogenic IHC?
Look first for staining outlining nuclei: NUP62 is a nuclear pore component, and the nuclear membrane is its main supported location (UniProt P37198: nuclear pore complex; HPA: nuclear membrane supported). HPA also reports cytoplasmic expression in several tissue cases, so cytoplasmic staining alone needs comparison with the expected nuclear envelope pattern and controls (HPA: tissue IHC profile). Nucleoplasmic localisation is additionally supported by cellular imaging, but its appearance in a chromogenic section may be difficult to separate from an envelope rim (HPA: subcellular profile; standard IHC practice). During mitosis, spindle pole association is documented; assess such staining with cell morphology rather than treating every cytoplasmic focus as a pore (UniProt P37198).
Could epitope position explain an unexpected NUP62 staining pattern?
Check the antibody's stated immunogen or epitope before attributing a staining difference to NUP62 biology (standard IHC practice). The record lists 0 annotated isoforms; that annotation does not establish which sequence this antibody recognises (UniProt P37198: isoforms; standard IHC practice). NUP62 has no annotated transmembrane segment, and the record lists glycosylation sites at residues 373 and 468 and phosphorylation at 408 and 418 (UniProt P37198: topology and modifications). Those features alone do not demonstrate epitope masking in this section; compare retrieval conditions and use independently validated staining evidence before assigning a modification-dependent explanation (standard IHC practice).
How can IF help resolve ambiguous NUP62 staining in tissue?
On a separately optimised IF section, examine whether NUP62 signal outlines nuclei, its main supported location, using a nuclear counterstain (HPA: nuclear membrane supported; standard IF practice). Multiplex with a marker identifying the expected cell population, such as glandular cells when assessing rectum, and establish each channel with single-label controls (HPA: high staining in rectal glandular cells; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and keep acquisition settings consistent across controls (standard IF practice). Because the antibody epitope's accessible side is unspecified and NUP62 has no transmembrane segment, optimise mild permeabilisation empirically rather than assuming an exposed membrane-facing epitope (UniProt P37198: topology; standard IF practice).
What should I check when NUP62 IHC shows diffuse brown background?
Compare the section with a no-primary control to assess background from secondary reagents, endogenous peroxidase and chromogen development (standard IHC practice). The selected tissue caption used 10% goat serum blocking, a peroxidase-conjugated secondary antibody and DAB; these report the tested detection setup, not a background-free result (caption A03950-2). Check peroxidase blocking, washing, primary antibody concentration and DAB development while keeping the EDTA retrieval at pH 8.0 consistent (standard IHC practice; datasheet A03950-2). Prioritise interpretable nuclear envelope staining over total brown intensity, because HPA describes nuclear membrane expression in most cell types and cytoplasmic expression in several cases (HPA: tissue IHC profile).
How should I quantify NUP62 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measuring NUP62; the nuclear membrane is its main supported location (HPA: nuclear membrane supported; standard IHC practice). For chromogenic sections, report the percentage of positive cells and an intensity-weighted H-score, or count positive cells per mm² when spatial density is the question (standard IHC practice). Normalise cell-based results to the number of evaluable cells in that population, and density results to analysed tissue area; exclude damaged and necrotic regions consistently (standard IHC practice). Keep retrieval, detection, counterstain, imaging and positivity thresholds matched, and report nuclear envelope and cytoplasmic scores separately when both are assessed (datasheet A03950-2: retrieval; HPA: tissue IHC profile; standard IHC practice).
When is a brown NUP62 signal likely to be artefactual?
A convincing result should include staining at the nuclear envelope in appropriate intact cells, consistent with NUP62's pore location and HPA tissue profile (UniProt P37198: nuclear pore complex; HPA: nuclear membrane expression). Strong staining restricted to cut edges, folds or necrotic areas warrants comparison with adjacent intact tissue and a no-primary control (standard IHC practice). Brown signal persisting without primary antibody suggests detection background, including endogenous peroxidase, rather than verified NUP62 staining (standard IHC practice). Cytoplasmic staining is possible in several tissue cases, and spindle pole association occurs during mitosis; interpret either pattern with morphology, expected cell population and control sections (HPA: tissue IHC profile; UniProt P37198: mitotic localisation; standard IHC practice).
Boster reagents
Best NUP62 / Nuclear pore glycoprotein p62 IHC Antibodies
Anti-NUP62 antibodies have IHC data from a human paraffin-embedded ovarian cancer section and IF data from human U2OS and HeLa cells; both list Human, Mouse and Rat reactivity (catalog images; catalog reactivity).
Real IHC data
Anti-NUP62 Antibody ®
Cat # A03950-2
Real IF data
Anti-NUP62 Rabbit Monoclonal Antibody
Cat # M03950
A03950-2 will render with IHC data from a human paraffin-embedded ovarian cancer section; its catalog also lists IF/ICC and shows IF in U2OS cells (A03950-2 image captions; catalog applications). M03950 will render with IF data from HeLa cells and lists IF/ICC applications (M03950 image caption; catalog applications).
Which to pick: Choose A03950-2 for tissue IHC: its image shows staining in a paraffin-embedded section after EDTA pH 8.0 heat retrieval at 2 μg/ml; the fixative is unreported (A03950-2 IHC image caption). For IF/ICC, both SKUs are listed, with A03950-2 shown in U2OS cells at 5 μg/ml and rabbit monoclonal M03950 shown in HeLa cells at 1:50 (catalog applications; A03950-2 and M03950 IF image captions; M03950 catalog title). Both list Human, Mouse and Rat reactivity, but the supplied IHC and IF images show human samples only (catalog reactivity; image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P37198 (NUP62_HUMAN, Nuclear pore glycoprotein p62).
- Human Protein Atlas. NUP62 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. NUP62 subcellular location (ICC-IF): Mainly localized to the nuclear membrane. In addition localized to the nucleoplasm..
- Human Protein Atlas. NUP62 antibody validation summary (2 antibodies).
- Traumatic injury compromises nucleocytoplasmic transport and leads to TDP-43 pathology. eLife 2021 — PMC8169113.
- Multiple lines of evidence for disruption of nuclear lamina and nucleoporins in FUS amyotrophic lateral sclerosis. Brain : a journal of neurology 2024 — PMC11684083.
- Mutant Huntingtin Disrupts the Nuclear Pore Complex. Neuron 2017 — PMC5595097.
- Exploring NUP62's role in cancer progression, tumor immunity, and treatment response: insights from multi-omics analysis. Frontiers in immunology 2025 — PMC11911477.
- PubMed PMID:1915414 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUP62 (Nuclear pore glycoprotein p62). Expected localisation Nuclear membrane in most cells; cytoplasm in some (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03950-2). Validated IHC antibody: A03950-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A03950-2); 2. Fixation: Image fixative and duration unreported (datasheet A03950-2); 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 A03950-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03950-2); 8. Primary antibody: Rabbit anti-NUP62, 2-5 μg/ml (datasheet A03950-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03950-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Parathyroid glandular cells show low staining (HPA tissue IHC); Isoform / epitope: No isoforms annotated; no transmembrane segment (UniProt); Regulation: No specific regulator annotated (UniProt). Evidence: UniProt P37198, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nuclear membrane. In addition localized to…, Datasheet A03950-2, PMC8169113, PMC11684083. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUP62/immunohistochemistry-ihc-guide.