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NUCKS1 / Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 · IHC design guide
Design Immunohistochemistry for NUCKS1
Plan NUCKS1 paraffin-section IHC around its ubiquitous nuclear pattern (HPA tissue IHC). Start with the catalog antibody at 2–5 μg/ml (datasheet A04997-3), and compare high-staining esophageal squamous cells with low-staining small-intestine glands (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUCKS1 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 staining across tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear signal in glandular, epithelial and other cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04997-3) | |
| Positive control | Adrenal gland+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). Selected-image fixative and duration unreported (datasheet A04997-3); verify before use. | |
| Caveat | Small-intestine glands and marrow cells show low staining (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unannotated (UniProt) |
Section 1
Recommended NUCKS1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with four published NUCKS1 tissue IHC methods (PMC10426139; PMC10393983; PMC4151634; PMC12174342).
| Sample | Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A04997-3) |
| Fixation | Image fixative and duration unreported (datasheet A04997-3); 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 A04997-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04997-3) |
| Primary antibody | Rabbit anti-NUCKS1, 2-5 μg/ml (datasheet A04997-3) |
| Primary incubation | Overnight at 4 °C (datasheet A04997-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04997-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUCKS1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A04997-3); citric buffer pH 6.0 is a published alternative (PMC4151634).
Section 2
What Is the Expected NUCKS1 Staining Pattern?
NUCKS1 should stain nuclei across many cell types in paraffin sections: HPA describes ubiquitous nuclear expression with Supported tissue IHC reliability (HPA: tissue IHC). A nuclear or chromosome-associated pattern fits its annotated location, and the protein has no transmembrane segment (UniProt Q9H1E3: location and topology). Compare intensity with the specific cell population shown by HPA; ubiquitous expression does not mean every cell must stain equally (HPA: tissue IHC).
| Distinct nuclear chromogen in glandular or epithelial cells, with limited cytoplasmic staining. | This fits the reported nuclear pattern (HPA: tissue IHC). HPA records High staining in adrenal glandular cells and bronchial respiratory epithelial cells; those are useful tissue and cell comparisons (HPA: High in adrenal gland and bronchus). |
| Predominantly cytoplasmic or membrane-like staining, with nuclei largely clear. | That compartment disagrees with nuclear and chromosome localization (UniProt Q9H1E3: location) and HPA's ubiquitous nuclear IHC profile (HPA: tissue IHC). Review staining and counterstain before interpreting the cytoplasmic signal as NUCKS1. |
| Strong staining in a cell population expected to be Low in the selected tissue. | High signal in small-intestinal glandular cells or bone-marrow hematopoietic cells conflicts with their Low HPA entries (HPA: Low in small intestine and bone marrow). Consider cross-reactivity or endogenous detection activity; a discrepancy alone cannot identify the cause. |
| Diffuse chromogen across tissue, without discernible nuclear boundaries. | The pattern cannot be scored confidently as nuclear NUCKS1 (HPA: tissue IHC). General IHC practice is to inspect background, blocking, detection chemistry and counterstain alongside a control section before changing the interpretation. |
| No nuclear signal in a section expected to show High staining. | An unstained adrenal glandular or bronchial respiratory epithelial population conflicts with the listed High patterns (HPA: High in adrenal gland and bronchus). Check tissue preservation and the staining run; HPA does not establish NUCKS1-specific fixation sensitivity. |
💡Expected NUCKS1 appearanceCall a result positive when chromogen is clearly nuclear in the expected cells, especially an HPA High population; diffuse, predominantly cytoplasmic or membrane-like signal is suspect (HPA: tissue IHC; UniProt Q9H1E3: location).
| Tissue and cell context | HPA reports ubiquitous nuclear expression but different staining levels: selected glandular and epithelial populations are High, while small-intestinal glandular and bone-marrow hematopoietic cells are Low (HPA: tissue IHC). Compare the named cell population, not the entire section. |
| IHC antibody evidence | HPA062351 has Supported IHC status; HPA063843 has no IHC status listed (HPA: antibodies). The tissue profile has Supported reliability, described as high consistency between staining and RNA data (HPA: tissue IHC). These labels do not validate every staining condition. |
| Protein location and structure | NUCKS1 is annotated in the nucleus and on chromosomes, has no transmembrane segment, and is a single annotated chain from residues 1–243 (UniProt Q9H1E3). These facts support a nuclear readout; they do not establish antigen-retrieval needs. |
| Isoforms and modifications | UniProt lists 2 isoforms and 23 modified residues (UniProt Q9H1E3). Their presence alone does not predict which forms an IHC antibody detects or whether retrieval changes staining; no epitope-specific evidence is supplied. |
| IF/ICC Q: Where should fluorescence appear? | A: Mainly in the nucleoplasm, with additional nucleolar signal (HPA: supported ICC-IF locations). This is an interpretation cue only; IF/ICC workflow belongs to its separate guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclei are clear, but cytoplasm or cell borders are strongly colored. | The compartment conflicts with the reported nuclear location (HPA: tissue IHC; UniProt Q9H1E3: location). | Check the nuclear counterstain and compare a known HPA High cell population in the same run (HPA: High in adrenal gland or bronchus). Review detection background before calling the signal specific. |
| Small-intestinal glandular or bone-marrow hematopoietic cells stain as strongly as a High comparator. | Those populations are listed as Low; cross-reactivity or endogenous detection activity is possible, but the pattern does not distinguish them (HPA: tissue IHC). | Compare the same cell types with a detection control and assess whether staining remains nuclear. Treat the unexpected intensity as unresolved until controls explain it. |
| Chromogen is widespread and nuclear edges are difficult to distinguish. | Diffuse background can obscure the nuclear pattern (HPA: tissue IHC). The source record does not identify a NUCKS1-specific background mechanism. | Use general IHC controls to assess nonspecific detection, blocking and chromogen development. Adjust the staining workflow only after checking that the counterstain and a positive tissue remain interpretable. |
| An expected High population has no visible nuclear staining. | This conflicts with HPA's High entry for that cell population (HPA: High in adrenal gland or bronchus); the record does not establish a fixation-related explanation. | Verify tissue identity, run controls, reagent performance and the chosen IHC antibody's status. HPA062351 is Supported for IHC, whereas HPA063843 has no IHC status listed (HPA: antibodies). |
| A weakly stained field is called negative because few cells are conspicuous. | NUCKS1 is broadly nuclear, yet HPA reports Low staining in some populations (HPA: tissue IHC). Weak staining in a Low population alone cannot validate a negative call. | Score nuclei within identified cell populations and compare a High reference population from the same run (HPA: tissue IHC). Record weak signal separately from absent signal. |
| Nucleolar emphasis in IF/ICC is treated as an IHC failure. | HPA supports nucleoplasmic localization with additional nucleolar localization in ICC-IF (HPA: subcellular). Fluorescence detail need not appear identically with chromogenic IHC. | For this IHC guide, judge whether the chromogenic result is predominantly nuclear (HPA: tissue IHC). Use the separate IF/ICC guide for fluorescence workflow and its finer localization readout. |
Experiment evidence
IHC & IF images showing NUCKS1 staining
A04997-3 · IHCIHC analysis of NUCKS1 using anti-NUCKS1 antibody (A04997-3). NUCKS1 was detected in a paraffin-embedded section of human esophageal squamou
A04997-3 · IHCIHC analysis of NUCKS1 using anti-NUCKS1 antibody (A04997-3). NUCKS1 was detected in a paraffin-embedded section of human liver cancer tissu
A04997-3 · IFIF analysis of NUCKS1 using anti-NUCKS1 antibody (A04997-3). NUCKS1 was detected in a paraffin-embedded section of rat brain tissue. Heat me
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000069275-NUCKS1/tissue/cerebral+cortex#imid_19114667
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000069275-NUCKS1/tissue/colon#imid_19114609
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000069275-NUCKS1/tissue/liver#imid_19114602
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000069275-NUCKS1/tissue/kidney#imid_19114611
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA062351; green = NUCKS1, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA062351; green = NUCKS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUCKS1 IHC & IF
🧪Run adrenal gland first: glandular cells should show nuclear staining (HPA: High in adrenal gland glandular cells; UniProt Q9H1E3: nucleus). HPA detects NUCKS1 in all 45 scored tissues, so there is no listed negative tissue; no-primary and isotype controls carry the negative comparison, while any erythrocytes present on the positive slide should lack nuclear staining because they have no nucleus (HPA: no negative tissue; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: NUCKS1 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 NUCKS1 in MCF-7, U2OS, Rh30, SH-SY5Y, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, a concentration-matched nonimmune rabbit IgG control matched to the primary antibody’s clonality, and a NUCKS1 knockout specimen if available (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection and compare any brown adrenal pigment with the no-primary section (caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A04997-3 paraffin-section caption does not state a fixative (caption: fixative unreported). That caption uses heat retrieval in EDTA at pH 8.0, supporting a retrieval starting point without establishing that retrieval is essential; frozen-section performance is unreported (caption: EDTA heat retrieval; standard IHC practice). IF/ICC may make nucleoplasmic and nucleolar localization easier to inspect with a nuclear counterstain, though tissue autofluorescence needs a no-primary check (HPA: supported nucleoplasmic and additional nucleolar localization; standard IF practice).
HPA tissue IHC evidence for NUCKS1
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: NUCKS1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced NUCKS1 IHC Tips
Troubleshoot chromogenic NUCKS1 staining in paraffin sections by checking nuclear localisation, retrieval conditions, controls and scoring consistency (UniProt Q9H1E3; HPA tissue IHC).
How should I adjust retrieval when NUCKS1 nuclear staining is weak?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04997-3). Keep heating and cooling conditions consistent across test and control sections, then compare nuclear signal at the same antibody concentration; the catalog image used 2 μg/ml overnight at 4°C (datasheet A04997-3; standard IHC practice). If signal remains weak, vary retrieval duration in a small, documented series while watching for tissue damage and increased background (standard IHC practice). Judge improvement by nuclear staining in intact cells, because NUCKS1 is nuclear and chromosomal and HPA reports ubiquitous nuclear expression (UniProt Q9H1E3; HPA tissue IHC).
Could fixation explain faint or uneven NUCKS1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report a fixative (datasheet A04997-3). Record the actual fixative, fixation interval and processing history for each specimen, and compare sections with matched handling before changing the antibody concentration (standard IHC practice). Maintain the documented EDTA pH 8.0 retrieval while checking whether uneven signal follows processing differences or the edges of a section (datasheet A04997-3; standard IHC practice). Use intact nuclei and a consistently processed positive control to assess the result; neither HPA tissue staining nor NUCKS1 topology establishes a fixation response (HPA tissue IHC; UniProt Q9H1E3; standard IHC practice).
What staining pattern should count as NUCKS1 positive in tissue?
Score convincing signal in intact nuclei: NUCKS1 is assigned to the nucleus and chromosomes, while HPA describes ubiquitous nuclear tissue staining (UniProt Q9H1E3; HPA tissue IHC). Nucleoplasmic staining is the main supported subcellular pattern, with additional nucleolar localisation, so those distributions can guide review of well-preserved cells (HPA subcellular). Compare DAB staining with the nuclear counterstain at the same magnification and exclude pigment or diffuse deposits outside identifiable nuclei (standard IHC practice). A prominent cell-surface pattern needs specificity checks because NUCKS1 has no annotated transmembrane segment; compartment alone cannot prove antibody specificity (UniProt Q9H1E3; standard IHC practice).
Could isoforms or phosphorylation alter epitope detection in paraffin sections?
NUCKS1 has 2 annotated isoforms and multiple modified residues, including phosphotyrosine and phosphoserine sites, but the supplied evidence does not identify this antibody’s epitope (UniProt Q9H1E3; datasheet A04997-3). Do not assign a weak section to one isoform or phosphorylation state from DAB intensity alone; compare matched sections under the same EDTA pH 8.0 retrieval first (UniProt Q9H1E3; datasheet A04997-3; standard IHC practice). If epitope or modification dependence matters, obtain antibody-specific epitope information and use an independently validated control (standard IHC practice). Report the result as NUCKS1 immunoreactivity unless those controls establish narrower specificity (standard IHC practice).
How can I investigate the same localisation by multiplex IF?
For the separate IF/ICC workflow, pair NUCKS1 with a validated marker for the expected cell population and a nuclear counterstain, then inspect overlap within individual nuclei (HPA tissue IHC; HPA subcellular; standard IF practice). Choose a fluorophore channel with low specimen autofluorescence and include single-label and unstained controls to assess bleed-through and background (standard IF practice). NUCKS1 is nuclear and has no transmembrane segment, so permeabilise cells or sections sufficiently to give the antibody access to its intracellular epitope (UniProt Q9H1E3; standard IF practice). HPA reports mainly nucleoplasmic signal with additional nucleolar localisation, but its IF images do not validate the selected IHC antibody for multiplex IF (HPA subcellular; datasheet A04997-3).
How do I reduce diffuse DAB background without losing nuclear signal?
First compare a no-primary control with the test section to separate detection-system deposits from antibody-associated staining (standard IHC practice). The catalog tissue image used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary before DAB development (datasheet A04997-3). If background tracks the primary antibody, titrate its concentration and shorten DAB development while checking that intact nuclei remain distinguishable (standard IHC practice; UniProt Q9H1E3). Include a peroxidase-blocking step and review tissue edges and damaged areas; these are general chromogenic IHC controls, not NUCKS1-specific conditions established by the caption (standard IHC practice; datasheet A04997-3).
How should I quantify NUCKS1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define a nuclear scoring rule before examining groups, because the supported tissue profile is broadly nuclear and a cytoplasmic area measurement would answer a different question (HPA tissue IHC; UniProt Q9H1E3; standard IHC practice). Record the percentage of positive nuclei and staining intensity, or calculate a nuclear H-score on a 0–300 scale using intensity categories 0–3 (standard IHC practice). Normalise counts to the number of intact evaluable nuclei within the same annotated cell population and report excluded necrotic or damaged regions (standard IHC practice). Apply identical retrieval, detection, counterstain and image-analysis thresholds across sections; HPA’s high and low examples are context, not a universal positivity cutoff (datasheet A04997-3; HPA tissue IHC; standard IHC practice).
Which apparent positives need the strongest artefact checks?
Treat crisp staining in intact nuclei as the expected pattern, while checking unexpected membrane-dominant or diffuse extracellular signal against NUCKS1’s nuclear location and lack of a transmembrane segment (UniProt Q9H1E3; HPA tissue IHC). Review the cell type before calling a region negative: HPA reports high staining in esophageal squamous epithelial cells but low staining in small-intestinal glandular cells and bone-marrow hematopoietic cells (HPA tissue IHC). Exclude necrotic areas, section-edge enhancement, pigment and deposits that do not align with counterstained nuclei (standard IHC practice). A no-primary control and peroxidase blocking help identify detection background and endogenous enzyme signal before interpreting DAB as NUCKS1 (standard IHC practice).
Boster reagents
Best NUCKS1 / Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 IHC Antibodies
A04997-3 has human paraffin-section IHC images and mouse and rat brain-section IF images (catalog image captions); its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-NUCKS1 Antibody ®
Cat # A04997-3
A04997-3 is listed for IHC and IF, with human paraffin-section IHC images from esophageal squamous carcinoma, liver cancer, lung cancer and rectum adenocarcinoma tissue (catalog applications and IHC image captions). Its IF images show paraffin-embedded mouse and rat brain sections (catalog IF image captions).
Which to pick: For tissue IHC, choose A04997-3: its human paraffin-section images use EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog IHC image captions); the fixative is unreported (catalog IHC image captions). For IF, A04997-3 has mouse and rat brain-section images at 5 μg/ml (catalog IF image captions); ICC validation is unreported (catalog applications and image captions). For cross-species planning, A04997-3 lists human, mouse and rat reactivity, while its IHC dilution listing specifies human and mouse and its IF images show mouse and rat (catalog reactivity, dilution listing and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H1E3 (NUCKS_HUMAN, Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1).
- Human Protein Atlas. NUCKS1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. NUCKS1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli..
- Human Protein Atlas. NUCKS1 antibody validation summary (2 antibodies).
- Roles of increased NUCKS1 expression in endometriosis. BMC women's health 2023 — PMC10426139.
- NUCKS1, a LINC00629-upregulated gene, facilitated osteosarcoma progression and metastasis by elevating asparagine synthesis. Cell death & disease 2023 — PMC10393983.
- Immunohistochemical study of nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 in invasive breast carcinoma of no special type. Experimental and therapeutic medicine 2014 — PMC4151634.
- NUCKS1 promotes invasion and metastasis of colorectal cancer by stabilizing HDAC2 and activating AKT. Oncogenesis 2025 — PMC12174342.
- PubMed PMID:11298763 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUCKS1 (Nuclear ubiquitous casein and cyclin-dependent kinase substrate 1). Expected localisation Nuclear staining across tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04997-3). Validated IHC antibody: A04997-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A04997-3); 2. Fixation: Image fixative and duration unreported (datasheet A04997-3); 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 A04997-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04997-3); 8. Primary antibody: Rabbit anti-NUCKS1, 2-5 μg/ml (datasheet A04997-3); 9. Primary incubation: Overnight at 4 °C (datasheet A04997-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A04997-3); verify before use.; Caveat: Small-intestine glands and marrow cells show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unannotated (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt Q9H1E3, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A04997-3, PMC10426139, PMC10393983. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUCKS1/immunohistochemistry-ihc-guide.