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NFIC / Nuclear factor 1 C-type · IHC design guide
Design Immunohistochemistry for NFIC
Plan NFIC IHC in paraffin sections around the nuclear staining seen in most tissues (HPA tissue IHC). Start the IHC-validated antibody at 0.5–1 µg/mL (datasheet A04154-2) and score staining by cell type, using HPA’s high-staining glandular cells as a reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NFIC 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissues, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04154-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Skeletal muscle RNA enrichment may not predict IHC intensity (HPA tissue RNA; HPA tissue IHC) | |
| Regulation | Skeletal muscle-enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | 6 isoforms; isoform-specific epitope coverage is unknown (UniProt; datasheet A04154-2) |
Section 1
Recommended NFIC IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A04154-2) with two published NFIC chromogenic IHC protocols (PMC7940473; PMC8415269).
| Sample | Paraffin-embedded human pancreatic cancer tissue; fixative not specified (datasheet A04154-2) |
| Fixation | Image fixative and duration unreported (datasheet A04154-2); 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 A04154-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04154-2) |
| Primary antibody | Rabbit anti-NFIC, 0.5-1μg/ml (datasheet A04154-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04154-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04154-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NFIC-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A04154-2); both published protocols used citrate retrieval (PMC7940473; PMC8415269).
Section 2
What Is the Expected NFIC Staining Pattern?
NFIC is a nuclear, DNA-binding protein with no transmembrane segment (UniProt P08651: location, function, topology). In paraffin-section IHC, expect nuclear staining across many tissues, including strong staining in the specified epithelial, glandular, glomerular and alveolar cell populations (HPA: nuclear expression in most tissues; listed High cells). The tissue IHC profile has Supported reliability, reflecting high consistency between staining and RNA expression (HPA: tissue reliability).
| Distinct nuclear staining in bronchial respiratory epithelium or duodenal Brunner glands. | This fits the expected compartment and a High-staining cell population (UniProt P08651: nucleus; HPA: High in bronchial respiratory epithelial cells and Brunner glands). Assess the named cells rather than treating all cells in either section as equivalent controls. |
| Predominantly cytoplasmic or membrane staining, with little nuclear signal. | That distribution conflicts with NFIC's annotated nuclear location and lack of a transmembrane segment (UniProt P08651: location, topology). Consider nonspecific staining or a detection artefact; compartment mismatch alone does not establish which step failed. |
| Strong signal in adipocytes or bladder urothelial cells. | Those populations are reported as Not detected (HPA: adipocytes; urothelial cells). Check morphology and cell identity, then consider cross-reactivity or endogenous detection activity as possible explanations (general IHC practice); nearby cells may have a different profile. |
| Diffuse colour across cells and surrounding tissue, obscuring nuclear boundaries. | This cannot be scored confidently as NFIC nuclear staining (UniProt P08651: nucleus; general IHC practice). Uneven blocking, excess detection reagent or inadequate washing can produce broad background (general IHC practice); compare the control section. |
| No nuclear signal in a verified positive cell population. | Absent staining in, for example, adrenal glandular cells conflicts with their reported High level (HPA: High in adrenal glandular cells). Review section identity and assay controls before concluding that NFIC is absent; a failed assay can also produce a negative slide (general IHC practice). |
💡Expected NFIC appearanceCall the result positive when discrete nuclear chromogen is prominent in the specified High-staining cells, such as bronchial respiratory epithelium (HPA: High; UniProt P08651: nucleus); isolated cytoplasmic colour or strong staining in adipocytes is suspect (UniProt P08651: location; HPA: adipocytes Not detected).
| Tissue and cell selection | HPA reports nuclear expression in most tissues, yet levels differ by cell population: kidney glomerular cells are High, while lymph-node germinal-center cells are Low and adipocytes are Not detected (HPA: tissue IHC). Select and score the actual named population. |
| IHC antibody evidence | HPA052625, CAB004300 and CAB004674 each have Supported IHC status (HPA: antibody validation). HPA063560 has Supported ICC status but no listed IHC status (HPA: antibody validation); ICC support alone does not establish performance in paraffin-section IHC. |
| Isoforms and epitope coverage | Six NFIC isoforms are listed, but the supplied record gives no antibody epitope or isoform-specific staining evidence (UniProt P08651: isoforms; supplied HPA antibody data). Do not interpret differences between cells as isoform-specific IHC staining on this evidence. |
| Processing and membrane association | NFIC has no annotated signal peptide, propeptide or transmembrane segment; the annotated chain spans residues 1–508 (UniProt P08651: processing, topology). An extracellular or membrane-rim pattern would need independent validation before assignment to NFIC. |
| IF/ICC: what pattern is expected? | HPA reports enhanced nucleoplasmic and supported nucleolar fibrillar-center localization by ICC-IF (HPA: subcellular profile). This provides a compartment check for IF/ICC images; the detailed IF/ICC workflow belongs in its own guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| A High-staining reference population has no nuclear signal. | The assay or tissue identification may have failed; absent colour alone cannot establish biological absence (HPA: High in the selected reference cells; general IHC practice). | Confirm the named cells on a counterstained section, check the detection controls, then review the catalog antibody's IHC-P instructions and the run's retrieval and dilution settings (general IHC practice). |
| Nuclei are weak but the whole section is coloured. | Broad background reduces nuclear contrast (UniProt P08651: nucleus; general IHC practice). Excess antibody or insufficient blocking or washing are possible workflow causes (general IHC practice). | Compare a negative reagent control, review blocking and washes, and titrate the primary antibody within its documented IHC-P conditions (general IHC practice). |
| Cytoplasm or cell borders stain more strongly than nuclei. | The distribution conflicts with the annotated nuclear location and absence of a transmembrane segment (UniProt P08651: location, topology). | Check a known High nuclear population in the same run, then review antibody specificity and detection controls before calling the staining NFIC (HPA: listed High cells; general IHC practice). |
| Adipocytes or bladder urothelium appear strongly positive. | These are reported as Not detected, so misidentified cells, cross-reactivity or endogenous detection activity are possibilities (HPA: negative populations; general IHC practice). | Verify the cell type, inspect the control section and compare with a listed High population under the same detection conditions (HPA: tissue IHC; general IHC practice). |
| Different tissues show different signal strengths. | The HPA profile is cell-specific: some listed populations are High, others Low or Not detected (HPA: tissue IHC). A tissue-wide average can conceal that distinction. | Record compartment and intensity by named cell population, using the same scoring criteria across sections (HPA: tissue IHC; general IHC practice). |
| ICC-IF shows puncta within nuclei, but IHC looks broadly nuclear. | HPA resolves nucleoplasm and nucleolar fibrillar centers in ICC-IF, while its tissue IHC profile describes nuclear expression (HPA: subcellular and tissue profiles). | Score the IHC result at the nuclear-compartment level; use the separate IF/ICC guide for finer subnuclear interpretation (HPA: subcellular and tissue profiles). |
Experiment evidence
IHC & IF images showing NFIC staining
A04154-2 · IHCIHC analysis of NFIC using anti-NFIC antibody (A04154-2). NFIC was detected in paraffin-embedded section of human pancreatic cancer tissue.
A04154-2 · IHCIHC analysis of NFIC using anti-NFIC antibody (A04154-2). NFIC was detected in paraffin-embedded section of mouse skeletal muscle tissue. He
A04154-2 · IFIF analysis of NFIC using anti-NFIC antibody (A04154-2). NFIC was detected in paraffin-embedded section of human intestinal cancer tissue. H
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141905-NFIC/tissue/cerebral+cortex#imid_17034899
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141905-NFIC/tissue/colon#imid_17034815
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141905-NFIC/tissue/liver#imid_17034808
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000141905-NFIC/tissue/kidney#imid_17034825
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052625; green = NFIC, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052625; green = NFIC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NFIC IHC & IF
🧪Run bronchus first and expect nuclear staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells; UniProt P08651: nucleus). Use adipose tissue as the negative tissue, where adipocytes are listed as not detected; any unstained non-epithelial cells on the bronchus slide should show counterstain without specific nuclear chromogen, but their NFIC-negative status must be verified rather than assumed (HPA: Not detected in adipocytes; HPA: bronchus row specifies respiratory epithelial cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NFIC in HeLa, MCF-7, U2OS, SiHa, U-251MG, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced), Nucleoli fibrillar center (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only slide, a concentration-matched nonimmune rabbit IgG control matched to the primary’s clonality if known, and an NFIC knockout biological control where available (caption: rabbit primary; standard IHC controls). Quench endogenous peroxidase and check for endogenous biotin background in bronchus when using the caption’s streptavidin–biotin/DAB detection method (caption: SABC with DAB; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04154-2 tissue-IHC caption does not state the fixative (caption: fixative not stated). The paraffin-section example used heat retrieval in EDTA at pH 8.0, but it does not establish that NFIC staining requires that retrieval method (caption: EDTA pH 8.0 retrieval). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC; bronchial endogenous peroxidase can create chromogenic background, so interpret epithelial nuclear staining against the controls (HPA: High in bronchus respiratory epithelial cells; standard IHC practice).
HPA tissue IHC evidence for NFIC
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 → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glands of Brunner | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
Section 3
Advanced NFIC IHC Tips
Troubleshoot NFIC staining in paraffin sections by checking retrieval, nuclear localisation, controls and scoring before interpreting chromogenic signal.
How should I adjust antigen retrieval when NFIC nuclear staining is weak?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A04154-2). The selected image used this condition with 1 µg/mL primary antibody overnight at 4°C; keep those conditions stable while assessing retrieval (datasheet A04154-2). If nuclear signal remains weak, compare shorter and longer EDTA heating intervals on adjacent sections, checking tissue preservation and a matched positive control (standard IHC practice; HPA tissue IHC: nuclear expression in most tissues). If EDTA optimisation fails, trial citrate at pH 6.0 as a fallback with matched detection settings; increased diffuse staining alone does not establish specificity (standard IHC practice).
Can fixation explain patchy or absent NFIC staining in paraffin sections?
The selected paraffin-section caption does not state a fixative, so NFIC-specific fixation sensitivity is unknown (datasheet A04154-2). Record each block's actual fixative and exposure time, then compare matched sections using EDTA at pH 8.0 and the same detection settings (standard IHC practice; datasheet A04154-2). For patchy nuclei, inspect preservation across the section and compare a positive control before increasing primary concentration, which can also raise background (standard IHC practice). Neither NFIC's nuclear location nor its annotated phosphorylation sites establishes a preferred fixative or predicts a target-specific fixation effect (UniProt P08651: subcellular location and modified residues).
Which cellular compartment should count as positive for NFIC IHC?
Score convincing nuclear staining: NFIC is annotated as nuclear, and tissue IHC shows nuclear expression in most tissues (UniProt P08651: subcellular location; HPA tissue IHC: profile). Subcellular imaging further places it in the nucleoplasm and nucleoli fibrillar center, so some nuclear variation can be biologically plausible (HPA subcellular: main location). Use a counterstain to verify that chromogen overlaps intact nuclei, and keep cytoplasmic haze out of the positive-cell count (standard IHC practice). For tissue controls, respiratory epithelial cells of the bronchus have high reported staining, whereas adipocytes have no detected staining; compare the relevant cell populations rather than whole-section colour (HPA tissue IHC: bronchus and adipose tissue).
Could an epitope issue explain divergent NFIC staining across samples?
NFIC has 6 annotated isoforms, so establish which sequence the antibody recognises before treating staining differences as changes in total NFIC (UniProt P08651: isoforms). The supplied caption documents tissue staining with A04154-2 but does not map its epitope or establish equal recognition of those isoforms (datasheet A04154-2). Check the antibody's immunogen or epitope documentation, then compare sections with the same EDTA pH 8.0 retrieval and development time (standard IHC practice; datasheet A04154-2). Annotated phosphorylation sites include Ser194, Ser294, Tyr300 and Ser302; without epitope mapping, their effect on antibody binding is unknown (UniProt P08651: modified residues).
How can IF help check an ambiguous NFIC IHC pattern?
Use IF as a localisation cross-check: expected NFIC signal is nuclear, with reported nucleoplasmic and nucleolar fibrillar-center localisation (UniProt P08651: subcellular location; HPA subcellular: main location). Multiplex with a marker for the expected cell population, such as an epithelial marker when examining bronchial respiratory epithelium, and check overlap with a nuclear counterstain (HPA tissue IHC: bronchus; standard IF practice). Choose a fluorophore away from strong tissue autofluorescence, using matched unstained and secondary-only controls to assess that background (standard IF practice). Because NFIC is nuclear and has no transmembrane segment, permeabilisation must permit antibody access to the nuclear epitope; optimise it against nuclear preservation (UniProt P08651: subcellular location and topology; standard IF practice).
What should I check when DAB obscures NFIC nuclear staining?
First separate true nuclear signal from diffuse chromogen by reviewing a no-primary section and a known positive section developed for the same time (standard IHC practice; UniProt P08651: nucleus). The selected workflow used 10% goat serum blocking, biotinylated secondary antibody, a streptavidin-biotin complex and DAB, so assess background at each detection step (datasheet A04154-2). Confirm adequate endogenous peroxidase blocking and consider endogenous biotin where relevant to this detection chemistry (standard IHC practice; datasheet A04154-2: detection method). If background persists, shorten chromogen development or titrate the primary below the caption's 1 µg/mL while retaining a nuclear positive control (standard IHC practice; datasheet A04154-2).
How should I quantify NFIC staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear staining only, because NFIC is nuclear and tissue expression varies among cell types (UniProt P08651: subcellular location; HPA tissue IHC: profile). Report the percentage of positive viable nuclei and, when intensity is reproducible, an H-score using 0–3 intensity categories and a 0–300 scale (standard IHC practice). For spatial comparisons, positive nuclei per mm² can supplement percentage positive, provided the analysed area and cell-selection rules are stated (standard IHC practice). Normalise counts to evaluable nuclei within the same defined compartment, and hold retrieval, antibody concentration and DAB development constant across compared sections (standard IHC practice; datasheet A04154-2: retrieval and detection).
How do I distinguish genuine NFIC positivity from artefact?
Prioritise staining confined to intact nuclei, consistent with NFIC's nuclear annotation and the reported tissue IHC pattern (UniProt P08651: subcellular location; HPA tissue IHC: profile). Check the right cells: bronchial respiratory epithelial cells have high reported staining, while adipocytes are reported as not detected; adjacent cell types need separate assessment (HPA tissue IHC: bronchus and adipose tissue). Discount isolated edge staining, necrotic regions and diffuse cytoplasmic colour when assigning positive nuclei (standard IHC practice). Compare no-primary and peroxidase-block controls if DAB appears outside cells, because endogenous enzyme activity can mimic chromogenic signal (standard IHC practice; datasheet A04154-2: DAB detection).
Boster reagents
Best NFIC / Nuclear factor 1 C-type IHC Antibodies
A04154-2 has IHC images of human pancreatic cancer, mouse skeletal muscle and rat pancreas muscle sections, plus IF images of human intestinal cancer and mouse skeletal muscle sections (catalog image captions).
Real IHC data
Anti-NFIC/CTF Antibody ®
Cat # A04154-2
A04154-2 is listed for IHC and shown on paraffin sections of human pancreatic cancer, mouse skeletal muscle and rat pancreas muscle tissue (catalog applications and IHC image captions). The same SKU is listed for IF and shown on paraffin sections of human intestinal cancer and mouse skeletal muscle tissue (catalog applications and IF image captions).
Which to pick: For tissue IHC, choose A04154-2 for paraffin sections; its IHC captions document EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml (catalog IHC image captions). For IF, A04154-2 has paraffin-section examples at 2 μg/ml; the payload does not document ICC (catalog IF image captions and applications). For cross-species IHC, its captions show human, mouse and rat sections; the fixative is unreported (catalog IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P08651 (NFIC_HUMAN, Nuclear factor 1 C-type).
- Human Protein Atlas. NFIC tissue IHC expression (reliability: Supported).
- Human Protein Atlas. NFIC subcellular location (ICC-IF): Localized to the nucleoplasm and nucleoli fibrillar center..
- Human Protein Atlas. NFIC antibody validation summary (4 antibodies).
- USP34 regulates tooth root morphogenesis by stabilizing NFIC. International journal of oral science 2021 — PMC7940473.
- Epithelial Bone Morphogenic Protein 2 and 4 Are Indispensable for Tooth Development. Frontiers in physiology 2021 — PMC8415269.
- Inactivation of Tgfbr2 in Osterix-Cre expressing dental mesenchyme disrupts molar root formation. Developmental biology 2013 — PMC3783640.
- Bone morphogenetic protein-2 gene controls tooth root development in coordination with formation of the periodontium. International journal of oral science 2013 — PMC3707077.
- PubMed PMID:3398920 — UniProt-cited evidence.
- PubMed PMID:8710515 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NFIC (Nuclear factor 1 C-type). Expected localisation Nuclear in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04154-2). Validated IHC antibody: A04154-2 at 0.5-1μg/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 pancreatic cancer tissue; fixative not specified (datasheet A04154-2); 2. Fixation: Image fixative and duration unreported (datasheet A04154-2); 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 A04154-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04154-2); 8. Primary antibody: Rabbit anti-NFIC, 0.5-1μg/ml (datasheet A04154-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04154-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Skeletal muscle RNA enrichment may not predict IHC intensity (HPA tissue RNA; HPA tissue IHC); Isoform / epitope: 6 isoforms; isoform-specific epitope coverage is unknown (UniProt; datasheet A04154-2); Regulation: Skeletal muscle-enhanced RNA (HPA tissue RNA). Evidence: UniProt P08651, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm and nucleoli fibrillar center., Datasheet A04154-2, PMC7940473, PMC8415269. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NFIC/immunohistochemistry-ihc-guide.