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NUDCD1 / NudC domain-containing protein 1 · IHC design guide
Design Immunohistochemistry for NUDCD1
For human paraffin sections, use the IHC-validated antibody at 2–5 μg/ml for chromogenic NUDCD1 staining (datasheet A10778-1). Assess cytoplasmic and nuclear staining, including high levels in smooth muscle cells, while accounting for uncertain tissue-IHC reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUDCD1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | High smooth muscle staining, cytoplasmic and nuclear (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10778-1) | |
| Positive control | Adrenal gland+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 conditions consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Isoform 2 is predominantly expressed in testis (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended NUDCD1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published NUDCD1 staining methods for colorectal tissue and paraffin sections (PMC9476325; PMC8351729).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A10778-1) |
| Fixation | Image fixative and duration unreported (datasheet A10778-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A10778-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10778-1) |
| Primary antibody | Rabbit anti-NUDCD1, 2-5 μg/ml (datasheet A10778-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10778-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10778-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUDCD1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues, high levels in smooth muscle cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A10778-1); citrate retrieval at pH 6.0 is a published alternative (PMC9476325).
Section 2
What Is the Expected NUDCD1 Staining Pattern?
NUDCD1 is reported in the cytoplasm and nucleus and has no transmembrane segment (UniProt Q96RS6 topology and subcellular location). In tissue IHC, expect staining in smooth muscle cells and in several glandular, glial, and kidney tubular cell populations (HPA: tissue IHC). Treat this as a provisional pattern: HPA rates the tissue IHC evidence Uncertain because antibody staining and RNA expression show low consistency (HPA: tissue reliability).
| Smooth muscle cells stain strongly, while selected glandular cells, glia, or kidney tubular cells show more moderate nuclear and cytoplasmic staining (HPA: tissue IHC). | This fits the reported tissue distribution and compartments (HPA: tissue IHC; UniProt Q96RS6 subcellular location). Record the cell type and compartment separately: a positive field should not be reduced to a single whole-section intensity score (general IHC practice). |
| The dominant signal outlines cell borders or appears confined to a membrane, with little intracellular staining. | A membrane-only pattern conflicts with the reported nuclear and cytoplasmic distribution and the absence of a transmembrane segment (UniProt Q96RS6 topology and subcellular location). Check morphology, detection controls, and antibody specificity before assigning it to NUDCD1 (general IHC practice). |
| Adipocytes, esophageal squamous epithelium, or prostatic glandular cells stain prominently. | HPA lists these specific cell populations as not detected by tissue IHC (HPA: adipose tissue, esophagus, prostate). Prominent staining warrants review for cross-reactivity or endogenous detection activity (general IHC practice); HPA's uncertain reliability prevents calling every such focus definitively false (HPA: tissue reliability). |
| Color spreads across tissue structures, blank areas, or multiple unrelated cell types without clear cellular boundaries. | This is difficult to interpret as the reported compartment-specific pattern (HPA: tissue IHC). Diffuse signal can arise from background staining or detection chemistry (general IHC practice); compare a no-primary control and assess whether staining follows identifiable cells (general IHC practice). |
| No staining appears in smooth muscle cells or in HPA-reported medium-level cell populations. | An absent signal in those populations conflicts with the reported IHC observations (HPA: tissue IHC), but it does not alone prove target absence. Review section quality and assay controls, then optimize general IHC conditions before interpreting a negative result (general IHC practice). |
💡Expected NUDCD1 appearanceA plausible positive result is identifiable nuclear and cytoplasmic staining, high in smooth muscle cells and medium in specified glandular, glial, or kidney tubular cells; isolated membrane outlines or widespread cell-free color should prompt a specificity or background check (HPA: tissue IHC; UniProt Q96RS6 topology; general IHC practice).
| Compartment and topology (UniProt Q96RS6 subcellular location and topology) | NUDCD1 is annotated in nucleus and cytoplasm, without a transmembrane segment (UniProt Q96RS6). Score intracellular signal by compartment; a membrane-only pattern needs independent verification (general IHC practice). |
| Cell-specific reference pattern (HPA: tissue IHC) | HPA reports high staining in smooth muscle cells and medium staining in listed glandular cells, glia, and kidney tubular cells (HPA: tissue IHC). Its listed negative cell populations are useful comparison regions, subject to the Uncertain reliability rating (HPA: tissue reliability). |
| Isoform expression (UniProt Q96RS6 tissue specificity) | Isoform 1 is reported in leukemias, various solid tumor cell lines, testis, and heart; isoform 2 is predominant in testis and weak in tumor cells (UniProt Q96RS6 tissue specificity). The supplied record does not establish which isoforms an IHC antibody detects; avoid isoform-specific conclusions from staining alone. |
| Antibody validation (HPA: HPA023183) | HPA labels this antibody's IHC result Uncertain, while its ICC result is Supported (HPA: HPA023183). Keep those application-specific assessments distinct when judging tissue sections; an ICC validation label does not resolve the tissue IHC uncertainty (HPA: antibody validation; tissue reliability). |
| Q: What should IF/ICC show? A: Mainly nucleoplasmic signal, with additional cytosolic signal (HPA: subcellular ICC-IF). | The supported ICC-IF localization is nucleoplasm plus cytosol (HPA: subcellular ICC-IF). Use it as a compartment comparison only; tissue IHC retains its separate Uncertain reliability rating (HPA: tissue reliability). |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive-looking field shows only membrane outlines. | The pattern conflicts with the annotated nuclear and cytoplasmic location and lack of a transmembrane segment (UniProt Q96RS6). | Inspect cellular morphology and compare a no-primary control; confirm the compartment with an independent specificity check before scoring NUDCD1 (general IHC practice). |
| Strong color appears in adipocytes, esophageal squamous cells, or prostatic glands. | Those cell populations are listed as not detected in HPA tissue IHC (HPA: adipose tissue, esophagus, prostate); cross-reactivity or endogenous activity is possible (general IHC practice). | Run a no-primary control and check whether the signal is cell-bound and reproducible (general IHC practice). Interpret discordance cautiously because HPA tissue reliability is uncertain (HPA: tissue reliability). |
| The entire section has diffuse brown background. | Nonspecific binding or endogenous enzyme activity can obscure cell-specific chromogenic staining (general IHC practice). | Compare no-primary and detection controls; review blocking, washes, and chromogen development, then reassess nuclear and cytoplasmic boundaries (general IHC practice; UniProt Q96RS6 subcellular location). |
| A section expected to contain smooth muscle cells has no detectable signal. | HPA reports high staining in smooth muscle cells, so a blank section may reflect assay performance or sampling (HPA: tissue IHC; general IHC practice). | Confirm that the relevant cells are present, inspect a positive control section, and review retrieval and detection settings using standard IHC controls (general IHC practice). Do not infer NUDCD1-specific fixation sensitivity from this result. |
| Only one intracellular compartment stains in tissue IHC. | HPA tissue IHC describes nuclear and cytoplasmic expression, but its evidence is uncertain (HPA: tissue profile and reliability); a single compartment is therefore a discrepancy to assess, not a definitive failure. | Score nucleus and cytoplasm separately, review counterstain and section morphology, and compare an independent antibody or orthogonal result if available (general IHC practice). |
| ICC-IF looks convincing, but the paraffin-section IHC pattern remains ambiguous. | The antibody has supported ICC and uncertain IHC validation, which are different application assessments (HPA: HPA023183). | Use the ICC-IF nucleoplasm and cytosol pattern as a localization reference (HPA: subcellular ICC-IF); resolve tissue IHC with tissue-specific controls and cautious scoring (general IHC practice; HPA: tissue reliability). |
Experiment evidence
IHC & IF images showing NUDCD1 staining
A10778-1 · IHCIHC analysis of NUDCD1 using anti-NUDCD1 antibody (A10778-1). NUDCD1 was detected in a paraffin-embedded section of human lung cancer tissue
A10778-1 · IHCIHC analysis of NUDCD1 using anti-NUDCD1 antibody (A10778-1). NUDCD1 was detected in a paraffin-embedded section of human ovarian cancer tis
A10778-1 · IFIF analysis of NUDCD1 using anti-NUDCD1 antibody (A10778-1) and anti-Beta Tubulin antibody (M01857-3). NUDCD1 was detected in immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120526-NUDCD1/tissue/cerebral+cortex#imid_6078936
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120526-NUDCD1/tissue/colon#imid_6078870
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120526-NUDCD1/tissue/liver#imid_6078863
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000120526-NUDCD1/tissue/kidney#imid_6078872
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA023183; green = NUDCD1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA023183; green = NUDCD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUDCD1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: Medium in colon glandular cells). Use esophagus squamous epithelium as the negative tissue (HPA: Not detected); neighboring colon cells count as internal negatives only where they remain unstained, since the supplied HPA row does not identify a negative colon cell type.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUDCD1 in A-431, U-251MG, NIH 3T3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus NUDCD1 knockout material if available or peptide competition when the immunogen is known (standard IHC practice). In colon, quench endogenous peroxidase and inspect inflammatory cells for background DAB signal (standard chromogenic IHC practice).
⚠️Feasibility: The selected A10778-1 paraffin-section caption does not report a fixative, and no target-specific fixation window or fixation effect is reported in the supplied evidence. That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for every specimen (selected A10778-1 tissue-IHC caption). HPA supports nucleoplasmic and cytosolic ICC-IF localization (HPA: subcellular), but the supplied evidence does not establish whether frozen sections or IF are easier than paraffin-section IHC.
HPA tissue IHC evidence for NUDCD1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUDCD1 IHC Tips
Troubleshoot NUDCD1 staining in paraffin sections by checking retrieval, compartment, cell type and assay controls before interpreting chromogenic signal.
Which retrieval conditions should I start with for NUDCD1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A10778-1). The supplied paraffin-section image used that retrieval before staining human lung cancer tissue with 2 μg/ml primary antibody overnight at 4°C (caption A10778-1). If staining is weak, compare retrieval duration on adjacent sections while holding antibody concentration and DAB development constant, and check tissue integrity after heating (standard IHC practice). Keep a no-primary control in each comparison so retrieval-related background is distinguishable from antibody-dependent staining (standard IHC practice). Record the chosen heating and cooling conditions for reproducible scoring (standard IHC practice).
How should I troubleshoot fixation-related loss of NUDCD1 staining?
The selected image describes a paraffin-embedded section but does not state its fixative, so NUDCD1-specific fixation sensitivity is unknown (caption A10778-1). Record the fixative and fixation duration for each specimen, then compare sections processed under documented conditions using the same EDTA pH 8.0 retrieval and staining run (datasheet A10778-1; standard IHC practice). Uneven staining across a section can reflect differences in tissue processing, so inspect morphology and include a matched control section before changing the primary antibody concentration (standard IHC practice). Do not infer a NUDCD1-specific fixation effect from its reported cytoplasmic and nuclear locations (UniProt Q96RS6).
Should NUDCD1 staining be nuclear, cytoplasmic or both?
Score nuclear and cytoplasmic staining separately: both compartments are reported for NUDCD1 (UniProt Q96RS6), while supported ICC/IF localisation is mainly nucleoplasmic with additional cytosolic signal (HPA subcellular). In paraffin sections, use a nuclear counterstain to define the nuclear boundary before assigning DAB to either compartment (standard IHC practice). Compare staining within the same cell type across cases, because the tissue IHC profile describes cytoplasmic and nuclear expression with high levels in smooth muscle cells (HPA tissue IHC). Treat a compartment pattern as supportive rather than decisive evidence, since tissue IHC reliability is rated uncertain owing to low consistency with RNA data (HPA tissue IHC).
Can this IHC stain distinguish NUDCD1 isoforms or epitope-dependent patterns?
NUDCD1 has 3 annotated isoforms, and the supplied antibody caption does not define an epitope or establish isoform discrimination (UniProt Q96RS6; caption A10778-1). Isoform 1 is reported in leukemias, various solid tumor cell lines, testis and heart, whereas isoform 2 is predominantly reported in testis and weakly in tumor cells (UniProt Q96RS6). Therefore, report this chromogenic signal as NUDCD1 immunoreactivity unless isoform specificity is established with independently documented reagents or orthogonal data (standard IHC interpretation). The annotated CS domain spans residues 273–361, but its presence alone does not identify the catalog antibody’s binding site (UniProt Q96RS6; caption A10778-1).
How should I assess NUDCD1 localisation by multiplex IF?
On the separate IF/ICC workflow, pair NUDCD1 with a marker for the cell population being assessed; a smooth muscle cell marker is useful where high expression is expected from the tissue IHC profile (HPA tissue IHC). Choose a fluorophore in a spectral channel with low measured tissue autofluorescence and include single-label controls before interpreting overlap (standard IF practice). NUDCD1 is reported in the nucleoplasm and cytosol and has no transmembrane segment, so use controlled permeabilisation to access its intracellular epitopes while checking that nuclear morphology remains intact (HPA subcellular; UniProt Q96RS6; standard IF practice). Interpret apparent colocalisation cautiously because the tissue IHC reliability assessment is uncertain (HPA tissue IHC).
What should I change when NUDCD1 DAB staining is diffuse?
First inspect a no-primary section for endogenous peroxidase activity and apply an appropriate peroxidase block before DAB development (standard chromogenic IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (caption A10778-1). If background remains high, compare shorter DAB development or a lower primary concentration on matched sections while retaining the same EDTA pH 8.0 retrieval (datasheet A10778-1; standard IHC practice). Check whether the diffuse signal follows tissue edges or damaged areas before counting it as cellular NUDCD1 staining (standard IHC practice).
How should I quantify NUDCD1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell type and region of interest before scoring, and record nuclear and cytoplasmic DAB separately because both locations are reported for NUDCD1 (UniProt Q96RS6). Within each compartment, use an H-score or percentage of positive cells with prespecified intensity thresholds; for spatial questions, report positive-cell density per mm² of viable tissue (standard IHC quantification practice). Normalise counts to the number of evaluable cells or viable tissue area, and apply identical thresholds, counterstaining and imaging settings across compared sections (standard IHC quantification practice). Report controls and uncertainty alongside the scores because the HPA tissue IHC assessment rates the staining-to-RNA agreement as uncertain (HPA tissue IHC).
How can I distinguish convincing NUDCD1 signal from artefact?
A plausible positive pattern is cellular nuclear or cytoplasmic staining, assessed in an identified cell population; NUDCD1 is reported in both compartments (UniProt Q96RS6). Supported ICC/IF data place it mainly in the nucleoplasm with additional cytosolic signal, while tissue IHC reports high levels in smooth muscle cells (HPA subcellular; HPA tissue IHC). Question signal confined to tissue edges, necrotic areas or unexpected extracellular deposits, and use a no-primary section to identify endogenous enzyme or detection background (standard IHC practice). Compare morphology, compartment and controls before calling positivity, since HPA rates its tissue staining reliability uncertain because of low agreement with RNA expression (HPA tissue IHC).
Boster reagents
Best NUDCD1 / NudC domain-containing protein 1 IHC Antibodies
A10778-1 has IHC images from human paraffin-embedded lung and ovarian cancer sections and an IF image from HELA cells (catalog: IHC/IF captions).
Real IHC data
Anti-NUDCD1 Antibody ®
Cat # A10778-1
A10778-1 will render with its IHC figure from a human paraffin-embedded lung cancer section (catalog: card figure caption). Its additional IHC caption documents human ovarian cancer tissue, and its IF caption documents HELA cells (catalog: IHC/IF captions).
Which to pick: Choose A10778-1 for human paraffin-section IHC: its caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog: IHC caption); the fixative is unreported (catalog: IHC caption). For IF/ICC, the same SKU has an IF image from HELA cells at 5 μg/ml (catalog: IF caption). Human, mouse and rat are listed as reactive species, but the supplied IHC and IF images document human samples only; clonality is unspecified (catalog: reactivity, IHC/IF captions, clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96RS6 (NUDC1_HUMAN, NudC domain-containing protein 1).
- Human Protein Atlas. NUDCD1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. NUDCD1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. NUDCD1 antibody validation summary (1 antibodies).
- Identifying the role of NUDCD1 in human tumors from clinical and molecular mechanisms: a study based on comprehensive bioinformatics and experimental validation. Aging 2023 — PMC10333089.
- NudCD1 as a prognostic marker in colorectal cancer and its role in the upregulation of cellular spindle assembly checkpoint genes and LIS1 pathways. BMC cancer 2022 — PMC9476325.
- NUDCD1 knockdown inhibits the proliferation, migration, and invasion of pancreatic cancer via the EMT process. Aging 2021 — PMC8351729.
- PubMed PMID:11416219 — UniProt-cited evidence.
- PubMed PMID:14688378 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUDCD1 (NudC domain-containing protein 1). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10778-1). Validated IHC antibody: A10778-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A10778-1); 2. Fixation: Image fixative and duration unreported (datasheet A10778-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A10778-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10778-1); 8. Primary antibody: Rabbit anti-NUDCD1, 2-5 μg/ml (datasheet A10778-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10778-1). Watch for: Fixation: Keep fixation conditions consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Isoform 2 is predominantly expressed in testis (UniProt). Evidence: UniProt Q96RS6, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A10778-1, PMC9476325, PMC8351729. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUDCD1/immunohistochemistry-ihc-guide.