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NCBP1 / Nuclear cap-binding protein subunit 1 · IHC design guide
Design Immunohistochemistry for NCBP1
Plan chromogenic IHC for NCBP1 in paraffin sections using its general nuclear tissue pattern (HPA tissue IHC). Start the catalog antibody at 2–5 μg/mL (datasheet A07424-3) and compare nuclear staining with low hepatocyte signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NCBP1 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 | General nuclear tissue staining (HPA tissue IHC); cytoplasm also annotated (UniProt) | |
| Staining pattern | Nuclear staining in glandular and respiratory epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07424-3) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Hepatocytes show low staining despite the broad nuclear pattern (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms or processing variants annotated; no transmembrane segment (UniProt) |
Section 1
Recommended NCBP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published NCBP1 protocols for lung tissue and xenograft tumors (PMC6787490) and lymphoid tissue (PMC10224985).
| Sample | Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A07424-3) |
| Fixation | Image fixative and duration unreported (datasheet A07424-3); 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 A07424-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07424-3) |
| Primary antibody | Rabbit anti-NCBP1, 2-5 μg/ml (datasheet A07424-3) |
| Primary incubation | Overnight at 4 °C (datasheet A07424-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07424-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NCBP1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A07424-3); the lung study reports 10 mM EDTA without specifying pH (PMC6787490).
Section 2
What Is the Expected NCBP1 Staining Pattern?
In paraffin-section IHC, expect general nuclear NCBP1 staining, including high staining in several glandular, respiratory epithelial, hematopoietic and neuronal cell populations (HPA: tissue IHC). HPA rates the tissue profile Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC). NCBP1 is also annotated in the cytoplasm and has no transmembrane segment (UniProt Q09161: localization and topology).
| Nuclei stain clearly in colon glandular cells, bronchial respiratory epithelial cells or cerebral cortex neurons (HPA: High in each). | This fits the general nuclear IHC profile (HPA: tissue IHC). Compare the stained cell population and compartment with adjacent cells; intensity alone cannot establish specificity (standard IHC interpretation). |
| The dominant signal outlines cell membranes or lies outside cells, without a convincing nuclear component. | Treat this as an artefact or unsupported localization in this IHC assay: NCBP1 has no transmembrane segment, and HPA describes general nuclear tissue staining (UniProt Q09161: topology; HPA: tissue IHC). |
| Soft-tissue fibroblasts stain strongly while expected positive cells on the same run stain weakly or not at all. | Question antibody specificity or endogenous detection activity before scoring the fibroblasts as positive; HPA reports fibroblasts as Not detected in soft tissue (HPA: tissue IHC). A single unexpected cell is not definitive evidence of cross-reactivity (standard IHC interpretation). |
| Color spreads across nuclei, cytoplasm and tissue spaces, obscuring cell boundaries. | This is diffuse background rather than an interpretable compartment pattern (standard IHC interpretation). Compare a no-primary control and check blocking, washes and detection conditions before assigning NCBP1 localization (standard IHC practice). |
| No convincing staining appears in a section expected to contain high-staining cells. | First verify that the relevant cells are present and preserved; HPA reports High staining in bone-marrow hematopoietic cells and adrenal glandular cells (HPA: tissue IHC). A failed positive control calls the run into question before a specimen is scored negative (standard IHC practice). |
💡Expected NCBP1 appearanceA credible positive result is cell-associated nuclear staining in an HPA high-staining population, such as colon glandular cells; strong membrane outlines or stain in empty tissue spaces should prompt an artefact check (HPA: tissue IHC; UniProt Q09161: topology; standard IHC interpretation).
| Tissue and cell context (HPA: tissue IHC) | High staining is reported in specified cells of adrenal gland, bone marrow, bronchus, cerebral cortex, cervix, colon, duodenum and gallbladder; hepatocytes and parathyroid glandular cells are Low, while soft-tissue fibroblasts are Not detected (HPA: tissue IHC). |
| Antibody evidence (HPA: antibody validation) | HPA042411 and HPA049031 each have Enhanced IHC validation (HPA: antibodies). This supports using the tissue profile as a reference, while HPA still describes only medium consistency between staining and RNA expression (HPA: tissue IHC). |
| Intracellular localization and processing (UniProt Q09161) | UniProt annotates nucleus, cytoplasm and cytoplasmic mRNP granules, with no signal peptide, propeptide or transmembrane segment (UniProt Q09161). These annotations give no basis for expecting cell-surface or extracellular staining. |
| Retrieval and detection checks (standard IHC practice) | Follow the IHC-validated antibody's established retrieval and detection instructions when available; use run controls to assess performance (standard IHC practice). The supplied HPA and UniProt records do not establish NCBP1-specific fixation sensitivity or retrieval requirements. |
| IF/ICC Q: Should its pattern match paraffin-section IHC? (HPA: subcellular and tissue profiles) | A: HPA reports mainly cytosolic signal with additional nucleoplasmic signal in ICC-IF, while its tissue IHC profile is generally nuclear (HPA: subcellular; HPA: tissue IHC). Interpret each application against its own evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining cells show no signal (HPA: tissue IHC). | The run may have failed, or the expected cell population may be absent from the section (standard IHC interpretation). | Inspect tissue morphology and repeat with a known positive section; then review the IHC-validated antibody's retrieval, dilution and detection instructions (standard IHC practice). |
| Signal is mostly membranous or extracellular. | The pattern conflicts with general nuclear tissue staining and the absence of a transmembrane segment (HPA: tissue IHC; UniProt Q09161: topology). | Check a no-primary control and inspect precipitate, tissue edges and detection background before interpreting the signal as NCBP1 (standard IHC practice). |
| Soft-tissue fibroblasts appear strongly positive. | This differs from HPA's Not detected result for that cell population; nonspecific binding or endogenous detection activity is possible (HPA: tissue IHC; standard IHC interpretation). | Compare the no-primary control, check cell identification and review blocking and detection conditions before assigning positivity (standard IHC practice). |
| Stain is diffuse across the section. | Background can obscure cellular localization and intensity differences (standard IHC interpretation). | Review reagent dilution, blocking, wash steps and detection development against the established IHC workflow; reassess only after boundaries become readable (standard IHC practice). |
| Hepatocytes or parathyroid glandular cells stain less than colon glandular cells. | That contrast agrees with HPA's Low versus High tissue-IHC levels and does not alone indicate assay failure (HPA: tissue IHC). | Score each identified cell population separately and confirm run quality in an HPA high-staining reference tissue (HPA: tissue IHC; standard IHC practice). |
| ICC-IF looks cytosolic, but paraffin-section IHC looks nuclear. | Those are the distinct application profiles reported by HPA (HPA: subcellular ICC-IF; HPA: tissue IHC). | Review each image using its application-specific HPA profile and controls; do not transfer the ICC-IF compartment expectation directly to tissue IHC (HPA: subcellular; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing NCBP1 staining
A07424-3 · IHCIHC analysis of NCBP1 using anti-NCBP1 antibody (A07424-3). NCBP1 was detected in a paraffin-embedded section of human ovarian cancer tissue
A07424-3 · IHCIHC analysis of NCBP1 using anti-NCBP1 antibody (A07424-3). NCBP1 was detected in a paraffin-embedded section of human lung cancer tissue. H
A07424-3 · IFIF analysis of NCBP1 using anti-NCBP1 antibody (A07424-3). NCBP1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen
A07424-1 · IFImmunocytochemistry of NCBP1 in HeLa cells with NCBP1 antibody at 2.5 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136937-NCBP1/tissue/cerebral+cortex#imid_13473611
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136937-NCBP1/tissue/colon#imid_13466409
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136937-NCBP1/tissue/liver#imid_13466402
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000136937-NCBP1/tissue/pancreas#imid_13466361
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042411; green = NCBP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042411; green = NCBP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NCBP1 IHC & IF
🧪Run adrenal gland first and expect staining in its glandular cells (HPA: High in adrenal gland glandular cells). Use soft tissue fibroblasts as the biological negative (HPA: Not detected in soft tissue fibroblasts); on the adrenal slide, cells without specific staining should remain at background, but the supplied HPA rows do not identify a particular internal cell type as negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NCBP1 in A-431, U-251MG, U2OS, Rh30, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and a concentration-matched nonimmune rabbit IgG isotype control; confirm specificity with NCBP1 knockout material processed in parallel or peptide competition if the immunizing peptide is available (caption: rabbit primary; standard IHC controls). Quench endogenous peroxidase for HRP/DAB detection and inspect adrenal pigment that could resemble brown chromogen (caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the exact A07424-3 tissue-IHC caption does not state a fixative (caption: fixative not stated). The caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required (caption: EDTA retrieval). Is IF/ICC or a frozen section easier? The supplied evidence does not establish either as easier; HPA reports supported cytosolic and nucleoplasmic ICC-IF localization, while adrenal pigment warrants care when interpreting brown DAB signal (HPA: subcellular; standard IHC practice).
HPA tissue IHC evidence for NCBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NCBP1 IHC Tips
Use the catalog antibody’s paraffin-section workflow as the IHC starting point, then assess NCBP1 staining by compartment, cell type and controls.
How should I retrieve NCBP1 antigen when staining paraffin sections?
Start with heat-mediated antigen retrieval in EDTA buffer at pH 8.0 for paraffin sections (datasheet A07424-3). The selected tissue-IHC example used this retrieval before staining with 2 μg/ml antibody overnight at 4°C (datasheet A07424-3). If staining is weak, compare retrieval duration on adjacent sections while keeping antibody concentration and chromogen development consistent (standard IHC practice). Check whether stronger retrieval improves staining in viable cells without damaging section morphology or increasing diffuse background (standard IHC practice). Record heating and cooling conditions so the comparison can be repeated (standard IHC practice).
Could fixation explain weak NCBP1 staining in paraffin sections?
Target-specific sensitivity to fixation is unknown: the selected NCBP1 caption identifies a paraffin-embedded section but does not report its fixative (datasheet A07424-3). Record each specimen’s fixative and fixation duration before comparing staining across blocks (standard IHC practice). Test sections with comparable processing together, using EDTA retrieval at pH 8.0 and the same antibody incubation conditions (datasheet A07424-3; standard IHC practice). If signal differs, assess morphology and retrieval controls before attributing the difference to fixation (standard IHC practice). HPA tissue patterns cannot establish a fixation effect for this antibody (HPA tissue IHC; datasheet A07424-3).
Should I score nuclear or cytoplasmic NCBP1 staining?
Score nuclear and cytoplasmic staining separately, rather than combining them into one intensity value (standard IHC practice). HPA describes general nuclear expression in tissue IHC, while UniProt places NCBP1 in the nucleus and cytoplasm, including cytoplasmic mRNP granules (HPA tissue IHC; UniProt Q09161 subcellular). HPA’s ICC/IF summary instead reports mainly cytosolic signal with additional nucleoplasmic signal, so compartment expectations depend on assay context (HPA subcellular). For each paraffin section, record the stained cell type, compartment and intensity alongside the counterstain (standard IHC practice). Review unexpected diffuse staining against a no-primary control before interpreting it as NCBP1 (standard IHC practice).
How can epitope uncertainty affect NCBP1 IHC interpretation?
The supplied record lists 0 annotated isoforms and one NCBP1 chain spanning residues 1–790 (UniProt Q09161 processing and isoforms). Its MIF4G domain spans residues 28–240, and reported modified residues include positions 7, 21, 22, 201 and 204 (UniProt Q09161 domains and modified residues). The catalog antibody’s epitope is unspecified here, so these features do not establish whether retrieval or modification changes antibody binding (datasheet A07424-3; UniProt Q09161). Compare adjacent sections under matched detection conditions if assessing retrieval sensitivity (standard IHC practice). Avoid assigning a compartment-specific staining difference to an isoform without independent evidence (UniProt Q09161 isoforms; standard IHC practice).
How should I assess NCBP1 by IF alongside chromogenic IHC?
Use the separate IF/ICC guide for its protocol; treat IF here as a compartment check alongside paraffin-section IHC (HPA subcellular; datasheet A07424-3). Multiplex NCBP1 with a marker identifying the expected cell type, and compare its signal with a nuclear counterstain before assigning cytosolic or nucleoplasmic staining (standard IF practice). Choose a spectrally separated fluorophore, preferably toward red or far-red when tissue autofluorescence is prominent, and include single-channel controls (standard IF practice). NCBP1 has no transmembrane segment, so assess permeabilisation for access to intracellular cytosolic and nuclear epitopes without assuming the antibody’s epitope position (UniProt Q09161 topology and subcellular; standard IF practice).
What should I check when NCBP1 DAB staining looks diffuse?
Run a no-primary control and inspect it for endogenous peroxidase activity or detection-reagent staining before judging NCBP1 signal (standard IHC practice). Include a peroxidase-blocking step in the chromogenic workflow; the selected caption reports DAB development but does not describe that block (datasheet A07424-3; standard IHC practice). The selected section was blocked with 10% goat serum and incubated with 2 μg/ml antibody overnight at 4°C (datasheet A07424-3). Compare background in viable tissue, section edges and damaged areas, then adjust blocking or washing consistently across test sections (standard IHC practice). Interpret staining only after the negative control is acceptably clean (standard IHC practice).
How should I quantify NCBP1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report nuclear and cytoplasmic NCBP1 results separately (standard IHC practice; UniProt Q09161 subcellular). For nuclear staining, record the percentage of viable cells at each intensity and calculate an H-score from 0–300, or report percentage positive using a prespecified threshold (standard IHC practice). If counting spatially distinct positive cells, report density per mm² of viable tissue rather than per entire section (standard IHC practice). Keep retrieval, imaging and DAB development consistent across specimens and exclude necrotic or folded regions (standard IHC practice). State the denominator and scoring threshold with every comparison (standard IHC practice).
How can I distinguish plausible NCBP1 staining from artefact?
Look for staining within intact cells and compare its compartment with the nuclear tissue-IHC pattern and reported cytoplasmic localisation (HPA tissue IHC; UniProt Q09161 subcellular). HPA reports high staining in several glandular and other cell populations, but no detection in soft-tissue fibroblasts; identify the actual cell type before interpreting a positive area (HPA tissue IHC). Treat staining concentrated at section edges, folds or necrotic regions as suspect and review the no-primary control (standard IHC practice). Brown signal retained in that control may reflect endogenous enzyme or detection background rather than antibody-dependent staining (standard IHC practice). Resolve doubtful results using matched sections and consistent scoring criteria (standard IHC practice).
Boster reagents
Best NCBP1 / Nuclear cap-binding protein subunit 1 IHC Antibodies
Anti-NCBP1 antibodies have paraffin-section IHC and cell-based IF/ICC images from human samples (catalog image captions); both list human, mouse, and rat reactivity (catalog reactivity).
Real IHC data
Anti-NCBP1 Antibody ®
Cat # A07424-3
Real IF data
Anti-NCBP1 Antibody
Cat # A07424-1
A07424-3 has IHC images from paraffin-embedded human ovarian, lung, colorectal adenocarcinoma, and liver cancer sections, plus IF in A549 cells (A07424-3 image captions). A07424-1 has an ICC image in HeLa cells and lists ICC among its applications (A07424-1 image caption; applications).
Which to pick: Choose A07424-3 for tissue IHC: its images show paraffin-embedded sections, and its IHC application is listed (A07424-3 IHC captions; applications); the fixative is unreported (A07424-3 IHC captions). For cell-based fluorescence, A07424-3 has an IF image in A549 cells, while A07424-1 has an ICC image in HeLa cells (respective image captions). Both list human, mouse, and rat reactivity, but the supplied IHC and IF/ICC 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 Q09161 (NCBP1_HUMAN, Nuclear cap-binding protein subunit 1).
- Human Protein Atlas. NCBP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NCBP1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. NCBP1 antibody validation summary (2 antibodies).
- NCBP1 promotes the development of lung adenocarcinoma through up-regulation of CUL4B. Journal of cellular and molecular medicine 2019 — PMC6787490.
- NCBP1 enhanced proliferation of DLBCL cells via METTL3-mediated m6A modification of c-Myc. Scientific reports 2023 — PMC10224985.
- PubMed PMID:8069914 — UniProt-cited evidence.
- PubMed PMID:7937105 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NCBP1 (Nuclear cap-binding protein subunit 1). Expected localisation General nuclear tissue staining (HPA tissue IHC); cytoplasm also annotated (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07424-3). Validated IHC antibody: A07424-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A07424-3); 2. Fixation: Image fixative and duration unreported (datasheet A07424-3); 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 A07424-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07424-3); 8. Primary antibody: Rabbit anti-NCBP1, 2-5 μg/ml (datasheet A07424-3); 9. Primary incubation: Overnight at 4 °C (datasheet A07424-3). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Hepatocytes show low staining despite the broad nuclear pattern (HPA tissue IHC); Isoform / epitope: No isoforms or processing variants annotated; no transmembrane segment (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q09161, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A07424-3, PMC6787490, PMC10224985. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NCBP1/immunohistochemistry-ihc-guide.