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FANCC / Fanconi anemia group C protein · IHC design guide
Design Immunohistochemistry for FANCC
Plan FANCC IHC-P around nuclear and cytoplasmic tissue staining, with high staining in testis spermatogonia (HPA tissue IHC). The IHC-validated antibody has a 1:50–1:100 dilution range (datasheet: A02387-1); this guide covers fixation, chromogenic detection, and interpretation.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
FANCC 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 and cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | High in spermatogonia; nuclear and cytoplasmic in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formalin-fixed paraffin testis is shown (selected-SKU IHC image A02387-1); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02387-1) | |
| Caveat | Staining and RNA levels show medium consistency (HPA tissue IHC) | |
| Regulation | Liver-enhanced RNA expression (HPA tissue RNA) | |
| Isoform / epitope | 0 annotated isoforms; no processing or mapped C-term epitope (UniProt; datasheet) |
Section 1
Recommended FANCC IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol with 2 published breast carcinoma IHC protocols (PMC2633285; PMC6439399).
| Sample | FFPE tissue sections (datasheet A02387-1; sample unspecified) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A02387-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-FANCC, 1:50-1:100 (datasheet A02387-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | FANCC-positive staining in spermatogonia cells of testis (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); assess nuclear staining (UniProt Q00597).
Section 2
What Is the Expected FANCC Staining Pattern?
FANCC is predominantly nuclear, with a smaller cytoplasmic pool (UniProt Q00597). IHC shows nuclear and cytoplasmic staining in most tissues (HPA tissue IHC: Approved; medium consistency with RNA). Expect the strongest listed signal in testis spermatogonia; several glandular, hematopoietic, epithelial, and neural cell populations show medium signal (HPA tissue IHC). FANCC has no transmembrane segment (UniProt Q00597 topology).
| Nuclear staining with some cytoplasmic staining in testis spermatogonia. | This fits the expected compartments and the highest listed cell specific signal (UniProt Q00597; HPA tissue IHC: High in spermatogonia). Score the cells and compartments separately; neighboring cells need not match their intensity (general IHC practice). |
| Predominantly nuclear staining in a medium signal reference, such as adrenal glandular cells or bone marrow hematopoietic cells. | A moderate result can support the expected pattern; FANCC need not appear uniformly strong across tissues (HPA tissue IHC: Medium in these cells; UniProt Q00597: major form nuclear). Compare like cell populations rather than whole section color (general IHC practice). |
| Strong membrane outlining or an exclusively cytoplasmic pattern across the positive cells. | Question the localization before calling it FANCC: the major form is nuclear, and FANCC lacks a transmembrane segment (UniProt Q00597). Check compartment assignment against the counterstain and assess antibody and detection controls (general IHC practice). |
| Prominent color in an HPA undetected cell population, such as adipocytes or lymph node germinal center cells. | This conflicts with the listed cell specific observations (HPA tissue IHC: Not detected in these cells). Cross reactivity or endogenous detection activity is possible; an HPA undetected call is a comparator, not proof that every cell in that tissue must be blank (general IHC practice). |
| Diffuse color across many compartments, or no staining in testis spermatogonia. | Diffuse color limits compartment scoring; testis spermatogonia are listed as High (HPA tissue IHC). Check background controls for the former and a known positive section for the latter before treating either result as FANCC biology (general IHC practice). |
💡Expected FANCC appearanceCall a section positive when appropriate cells show chiefly nuclear staining with possible weaker cytoplasmic staining, including strong spermatogonial signal in testis (UniProt Q00597; HPA tissue IHC: High in spermatogonia); dominant membrane outlines or diffuse, cell indiscriminate color warrant review (general IHC practice).
| Biological compartment | The major FANCC form is nuclear and the minor form cytoplasmic (UniProt Q00597); HPA reports both compartments in most tissues (HPA tissue IHC). Use a nuclear counterstain to judge whether chromogen overlaps nuclei (general IHC practice). |
| Choice of positive and comparison cells | Testis spermatogonia are High; adrenal glandular cells and bone marrow hematopoietic cells are Medium (HPA tissue IHC). Adipocytes, lymph node germinal center cells, and spleen red pulp cells are listed as Not detected (HPA tissue IHC). |
| Strength of the reference evidence | HPA rates the tissue IHC profile Approved, with medium consistency between staining and RNA (HPA tissue IHC). Its antibody list marks CAB017793 IHC Approved; it does not assign an IHC rating to HPA030771 (HPA antibodies). |
| Protein architecture | FANCC has no transmembrane segment, signal peptide, propeptide, annotated isoforms, or glycosylation sites (UniProt Q00597). These annotations support checking a membrane dominant result, but do not establish an epitope retrieval requirement or fixation sensitivity. |
| IF/ICC Q: What location should I expect? | A: HPA supports nucleoplasmic localization in ICC IF images from A-431, U-251MG, and U2OS (HPA subcellular ICC IF). This is a localization cross check; the tissue IHC profile remains the reference for interpreting paraffin section chromogen (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A known positive testis section has no visible spermatogonial signal. | The result conflicts with the listed High cell specific staining (HPA tissue IHC); weak detection, unsuitable retrieval, or a staining run failure are possible (general IHC practice). | Check section integrity, reagent controls, and the IHC validated antibody's documented retrieval and dilution settings; repeat alongside a working positive control (general IHC practice). |
| Signal covers nuclei, cytoplasm, and empty spaces as a uniform haze. | Nonspecific background can obscure the expected nuclear dominant distribution (UniProt Q00597; general IHC practice). | Compare a no primary control, assess blocking and wash steps, and adjust detection exposure within the validated IHC workflow (general IHC practice). |
| Brown color appears in expected negative cells or persists without primary antibody. | Endogenous chromogen generating activity or nonspecific detection may contribute (general IHC practice). HPA lists adipocytes and germinal center cells as Not detected, cell specifically (HPA tissue IHC). | Use a no primary control and appropriate endogenous activity blocking for the detection chemistry; assess those cells separately from other cells in the section (general IHC practice). |
| A medium signal tissue looks negative while testis stains well. | The expected level differs by cell type: testis spermatogonia are High, while adrenal glandular and bone marrow hematopoietic cells are Medium (HPA tissue IHC). | Inspect the specified cells at comparable development and counterstain settings before changing the run; treat a faint result as uncertain if compartment and controls cannot be resolved (general IHC practice). |
| Color forms strong cell borders with little nuclear staining. | This is discordant with the nuclear major form and no transmembrane segment (UniProt Q00597); nonspecific staining or misread cell boundaries are possible (general IHC practice). | Check nuclear counterstain alignment and control sections, then review antibody specificity before scoring the border signal as FANCC (general IHC practice). |
| IHC and ICC IF seem to give different amounts of cytoplasmic staining. | HPA tissue IHC reports nuclear and cytoplasmic expression in most tissues, whereas its ICC IF summary supports nucleoplasm (HPA tissue IHC; HPA subcellular ICC IF). | Compare localization within each application and its controls; do not use ICC IF intensity to set a paraffin section positivity threshold (general IHC practice). |
Experiment evidence
IHC & IF images showing FANCC staining
A02387-1 · IHCFANCC Antibody (C-term) IHC analysis in formalin fixed and paraffin embedded testis tissue followed by peroxidase conjugation of the seconda
A02387-1 · IFImmunofluorescent analysis of U251 cells, using FANCC Antibody (C-term). A02387-1 was diluted at 1:25 dilution. Alexa Fluor 488-conjugated g
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000158169-FANCC/tissue/cerebral+cortex#imid_4560720
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000158169-FANCC/tissue/colon#imid_4560646
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000158169-FANCC/tissue/liver#imid_4560655
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000158169-FANCC/tissue/kidney#imid_4560656
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA030771; green = FANCC, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA030771; green = FANCC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for FANCC IHC & IF
🧪Run testis first and require staining in spermatogonia (High; HPA: testis, spermatogonia). Use adipose tissue as a negative comparator, scoring adipocytes, which are not detected (HPA: adipose tissue, adipocytes); on the testis slide, cells lacking specific signal should show counterstain only, but no other cell type is established as FANCC-negative by the supplied HPA row.
Positive control tissue: Testis (Spermatogonia cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show FANCC in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls matched to the primary antibody’s host species and clonality, plus FANCC knockout material where available or a peptide-block control if the immunizing peptide is available (standard IHC controls). For DAB detection in testis, block endogenous peroxidase and check the control slide for residual background (selected A02387-1 tissue-IHC caption: peroxidase secondary and DAB).
⚠️Feasibility: The selected A02387-1 testis caption explicitly reports formalin-fixed, paraffin-embedded tissue and DAB staining, but reports no target-specific fixation window or fixation effect (selected A02387-1 tissue-IHC caption). FANCC-specific antigen retrieval requirements are unreported; optimize retrieval for the catalog antibody before scoring (supplied target/application evidence). Frozen sections or IF cannot be judged easier from these data; HPA supports nucleoplasmic ICC-IF localization, while endogenous peroxidase is a potential DAB background source in the testis assay (HPA: subcellular; selected A02387-1 tissue-IHC caption; standard IHC practice).
HPA tissue IHC evidence for FANCC
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced FANCC IHC Tips
Troubleshoot FANCC staining in paraffin sections by checking retrieval, compartment, controls, and cell specific scoring (UniProt Q00597; HPA tissue IHC).
How should I retrieve FANCC in paraffin sections with weak nuclear staining?
Use Tris-EDTA at pH 9.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Let sections cool in retrieval buffer, then compare nuclear staining and tissue integrity across matched sections; the major FANCC form is nuclear (standard IHC practice; UniProt Q00597: major form nuclear). If staining remains weak, compare a citrate buffer retrieval condition on a separate section as an optimisation, without treating it as a validated FANCC condition (standard IHC practice). Include a no primary control and assess spermatogonia in testis as a positive reference, since HPA reports high staining there (standard IHC practice; HPA: High in testis spermatogonia).
Could fixation explain weak FANCC staining in my paraffin sections?
FANCC specific sensitivity to fixation duration is unknown from the supplied evidence, so record the fixative and processing history before changing conditions (supplied evidence; standard IHC practice). For poorly stained sections, compare blocks with documented fixation and run them through the same pH 9.0, 20 min retrieval and detection workflow (page retrieval rule; standard IHC practice). Check whether tissue morphology is preserved and whether a no primary control shows staining before attributing signal loss to fixation (standard IHC practice). Score nuclear and cytoplasmic staining separately because both locations are reported, with the major FANCC form nuclear (UniProt Q00597; HPA tissue IHC).
Should FANCC stain nuclei, cytoplasm, or both in tissue IHC?
Expect predominantly nuclear staining with possible cytoplasmic staining, because UniProt describes a major nuclear and minor cytoplasmic form and HPA reports both in tissues (UniProt Q00597; HPA tissue IHC). HPA subcellular imaging supports nucleoplasmic localisation, so assess whether nuclear signal lies within intact cell boundaries on the counterstained section (HPA subcellular; standard IHC practice). FANCC has no annotated transmembrane segment or signal peptide; a crisp membrane only pattern deserves a specificity check (UniProt Q00597 topology and processing; standard IHC practice). Compare nuclear and cytoplasmic compartments separately against a no primary control and a documented positive tissue section (standard IHC practice; HPA: High in testis spermatogonia).
How can epitope choice affect FANCC IHC interpretation?
The selected catalog antibody is described as C terminal and has a paraffin section image from formalin fixed testis with peroxidase secondary detection and DAB (SKU A02387-1 tissue IHC caption). UniProt lists a 558 amino acid chain and no annotated isoforms, glycosylation sites, or modified residues; those annotations do not establish antibody specificity (UniProt Q00597 processing, isoforms and PTMs; standard IHC practice). If staining changes after retrieval, compare matched sections and controls before assigning the change to epitope exposure (standard IHC practice). Interpret a positive result as staining with this antibody under these conditions, and use an independent specificity control when the biological conclusion depends on FANCC identity (standard IHC practice).
How should I adapt FANCC localisation checks for multiplex IF?
For a separate IF experiment, pair FANCC with a marker identifying the expected cell population; testis spermatogonia provide a relevant IHC reference population (HPA: High in testis spermatogonia; standard IF practice). Choose spectrally separated fluorophores after checking autofluorescence in an unstained section, and include single colour controls to assess bleed through (standard IF practice). FANCC has no transmembrane segment and is reported mainly in the nucleus, so use a permeabilisation step compatible with intracellular antigen access and check nuclear morphology (UniProt Q00597 topology and localisation; standard IF practice). The SKU A02387-1 caption documents chromogenic tissue IHC, so optimise IF staining and controls independently for the intended specimen (SKU A02387-1 tissue IHC caption; standard IF practice).
What should I check when FANCC DAB staining is diffuse or widespread?
Run a no primary section through the same peroxidase secondary and DAB steps to identify background from the detection workflow (standard IHC practice; SKU A02387-1 tissue IHC caption). Quench endogenous peroxidase, optimise blocking and washes, and compare antibody titrations on adjacent sections while keeping retrieval fixed at pH 9.0 for 20 min (standard IHC practice; page retrieval rule). FANCC is reported in most tissues, so widespread staining alone cannot establish background; inspect cellular boundaries and nuclear versus cytoplasmic distribution (HPA tissue IHC; UniProt Q00597 localisation; standard IHC practice). Exclude tissue edges, folds, necrotic areas and residual precipitate when judging whether diffuse DAB represents cellular signal (standard IHC practice).
How should I score heterogeneous FANCC staining across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear compartment before scoring, because FANCC is mainly nuclear and HPA reports different staining levels among cell types (UniProt Q00597 localisation; HPA tissue IHC). Record percent positive cells and intensity in consistent categories, then calculate a nuclear H score if the study requires one (standard IHC practice). Normalise positive counts to the number of evaluable cells in that population, or report positive cell density per mm² of evaluable tissue when counting cells is impractical (standard IHC practice). Score cytoplasmic signal separately and apply the same threshold, counterstain and imaging settings across compared sections (HPA tissue IHC; standard IHC practice).
How can I distinguish convincing FANCC staining from a DAB artefact?
Prioritise staining in intact nuclei with plausible cytoplasmic signal, consistent with the reported FANCC distribution and supported nucleoplasmic localisation (UniProt Q00597 localisation; HPA tissue IHC and subcellular). In testis, high staining in spermatogonia is a relevant reference, while signal assigned only to an unexpected cell population needs closer review (HPA: High in testis spermatogonia; standard IHC practice). Compare suspicious regions with nearby intact tissue and a no primary control; edge accentuation, necrotic debris and endogenous peroxidase can mimic cellular DAB signal (standard IHC practice). HPA rates tissue IHC reliability as Approved with medium RNA concordance, so resolve consequential claims with an independent specificity control (HPA tissue IHC; standard IHC practice).
Boster reagents
Best FANCC / Fanconi anemia group C protein IHC Antibodies
The human-reactive anti-FANCC antibody has IHC data from formalin-fixed, paraffin-embedded testis and IF data from U251 cells (catalog: human reactivity; IHC and IF image captions).
Real IHC data
Anti-FANCC Antibody (C-term)
Cat # A02387-1
A02387-1 will render with an IHC-P figure showing DAB staining of formalin-fixed, paraffin-embedded testis (catalog: A02387-1 IHC image caption). The same SKU is listed for human IF and has an IF image of U251 cells (catalog: A02387-1 applications, reactivity, and IF image caption).
Which to pick: Choose A02387-1 for human tissue IHC-P: its own figure documents formalin-fixed, paraffin-embedded testis, and its listed IHC-P dilution is 1:50–1:100 (catalog: A02387-1 IHC image caption and dilution). For IF/ICC, the same rabbit polyclonal antibody has an IF image of U251 cells at 1:25 (catalog: A02387-1 host, dilution_raw, and IF image caption). No cross-species choice is supported by this catalog; A02387-1 lists human reactivity only (catalog: A02387-1 reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q00597 (FANCC_HUMAN, Fanconi anemia group C protein).
- Human Protein Atlas. FANCC tissue IHC expression (reliability: Approved).
- Human Protein Atlas. FANCC subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. FANCC antibody validation summary (2 antibodies).
- Alterations in candidate genes PHF2, FANCC, PTCH1 and XPA at chromosomal 9q22.3 region: pathological significance in early- and late-onset breast carcinoma. Molecular cancer 2008 — PMC2633285.
- Deleterious Mutations in DNA Repair Gene FANCC Exist in BRCA1/2-Negative Chinese Familial Breast and/or Ovarian Cancer Patients. Frontiers in oncology 2019 — PMC6439399.
- Measuring PI3K Activation: Clinicopathologic, Immunohistochemical, and RNA Expression Analysis in Prostate Cancer. Molecular cancer research : MCR 2015 — PMC4618038.
- Zika virus depletes neural stem cells and evades selective autophagy by suppressing the Fanconi anemia protein FANCC. EMBO reports 2020 — PMC7726779.
- PubMed PMID:1574115 — UniProt-cited evidence.
- PubMed PMID:1641028 — UniProt-cited evidence.
- PubMed PMID:8490620 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for FANCC (Fanconi anemia group C protein). Expected localisation Nuclear and cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A02387-1 at 1:50-1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Spermatogonia cells); negative Adipose tissue. IHC-P workflow on FFPE sections. Protocol steps: 1. Sample: FFPE tissue sections (datasheet A02387-1; sample unspecified); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A02387-1); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-FANCC, 1:50-1:100 (datasheet A02387-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Formalin-fixed paraffin testis is shown (selected-SKU IHC image A02387-1); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02387-1); Caveat: Staining and RNA levels show medium consistency (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; no processing or mapped C-term epitope (UniProt; datasheet); Regulation: Liver-enhanced RNA expression (HPA tissue RNA). Evidence: UniProt Q00597, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A02387-1, PMC2633285, PMC6439399. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/FANCC/immunohistochemistry-ihc-guide.