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FSCN1 / Fascin · IHC design guide
Design Immunohistochemistry for FSCN1
Plan chromogenic IHC for FSCN1 using the catalog antibody at 1:50 on paraffin sections (datasheet M02147-1). Use cytoplasmic staining in the documented cell populations to guide interpretation (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
FSCN1 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 staining in tissue cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in tubules, glia, endothelium, fibroblasts and macrophages (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02147-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Bronchus+4 more · see all |
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 | Macrophage and endothelial staining can complicate cell scoring (HPA tissue IHC) | |
| Regulation | Low tissue specificity; staining varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | No isoforms annotated; Fascin chain spans residues 2–493 (UniProt) |
Section 1
Recommended FSCN1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M02147-1) is paired with four published FSCN1 IHC protocols (PMC2833258; PMC13243165; PMC13461984; PMC6494834).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet M02147-1) |
| Fixation | Image fixative and duration unreported (datasheet M02147-1); 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 M02147-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02147-1) |
| Primary antibody | Rabbit monoclonal (clone ADOA-6) anti-FSCN1, 1:50 (datasheet M02147-1) |
| Primary incubation | Overnight at 4 °C (datasheet M02147-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02147-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | FSCN1-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in cells in seminiferous tubules, glial cells, endothelial cells, fibroblasts and macrophages. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet M02147-1); the sarcoma study also used EDTA but did not report its pH (PMC13243165).
Section 2
What Is the Expected FSCN1 Staining Pattern?
FSCN1 should stain predominantly in the cytoplasm of selected cells, including glial cells, endothelial cells, fibroblasts, macrophages and cells in seminiferous tubules (HPA: tissue IHC profile). Cortical or projection-associated signal is also plausible because FSCN1 is an actin-bundling protein with no transmembrane segment (UniProt Q16658: function, localization and topology). The tissue IHC profile is rated Enhanced (HPA: reliability).
| Cytoplasmic staining is strong in lung endothelial cells, cerebral cortex glial cells or testis Sertoli cells, with adjacent cells showing different intensities (HPA: High in these cell populations). | This fits the cell-specific IHC pattern. Score the identified cells and their compartment rather than treating the whole tissue as uniformly positive; HPA reports staining by cell population (HPA: tissue IHC). |
| The apparent positive signal is confined to nuclei, with no convincing cytoplasmic staining in expected positive cells. | Treat an isolated nuclear pattern as suspect: UniProt places FSCN1 in the cytosol, cortex, cytoskeleton and projections, while HPA describes cytoplasmic tissue staining (UniProt Q16658: localization; HPA: tissue IHC profile). Check morphology and staining controls before scoring it. |
| Strong signal appears in bronchial respiratory epithelium or duodenal glandular cells (HPA: Not detected in these cell populations). | That distribution conflicts with the cited HPA observations, so investigate cross-reactivity or endogenous detection activity (HPA: tissue IHC; general IHC practice). It does not make every cell in the bronchus or duodenum a negative control. |
| A broad, poorly localized chromogenic haze covers positive and expected negative cell populations. | This is difficult to assign to FSCN1 because the reference pattern is cell-selective and mainly cytoplasmic (HPA: tissue IHC profile). Review blocking, antibody concentration and detection controls as general IHC troubleshooting steps (general IHC practice). |
| Known positive cells, such as lung endothelial cells or testis Sertoli cells, show no discernible cytoplasmic signal (HPA: High in these cells). | A failed positive control makes a negative result in the study specimen uninterpretable. Check tissue preservation, antigen retrieval and detection performance as general IHC workflow variables; the supplied sources do not define FSCN1-specific fixation sensitivity (general IHC practice). |
💡Expected FSCN1 appearanceCall a result positive when selected HPA-positive cells show clear, often strong cytoplasmic staining with plausible cortical or projection-associated detail; isolated nuclear staining or uniform staining of HPA-undetected cell populations warrants review (HPA: tissue IHC; UniProt Q16658: localization).
| Cell context and scoring | HPA reports High staining in breast glandular cells but Not detected in duodenal glandular cells; cell identity and tissue context therefore matter when choosing comparison fields (HPA: tissue IHC). Its low tissue RNA specificity is not a claim that every cell stains equally (HPA: RNA specificity and tissue IHC). |
| Target location and topology | FSCN1 has no transmembrane segment or signal peptide, and UniProt lists cytosolic, cortical, cytoskeletal and projection locations (UniProt Q16658: topology, processing and localization). Judge a chromogenic pattern against the identified cell compartment; a crisp surface outline alone is insufficient evidence of the expected tissue IHC pattern (HPA: cytoplasmic profile). |
| Strength of the reference pattern | The HPA tissue profile is Enhanced, with high consistency between staining and RNA data and support from similar staining by paired antibodies (HPA: reliability description). This supports the reference distribution but does not establish the performance of every antibody or preparation in a new laboratory. |
| IF/ICC Q&A: what location should be expected? | Expect mainly cytosol; HPA also lists plasma membrane with uncertain support in ICC-IF images (HPA: subcellular summary and location). UniProt additionally places FSCN1 at the cell cortex and actin-rich projections (UniProt Q16658: localization). This is an interpretation cue for IF/ICC, not an IHC-P protocol option. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in both study tissue and an HPA-positive control population. | The IHC run may have failed; absence in lung endothelial cells or testis Sertoli cells conflicts with the HPA reference pattern (HPA: High in these cells). | Check the control section, retrieval conditions, antibody dilution and detection reagents before interpreting the study tissue (general IHC practice). The supplied sources give no FSCN1-specific retrieval condition or fixation effect. |
| Background obscures cell boundaries across the section. | Nonspecific antibody binding or detection background can obscure a cell-selective cytoplasmic pattern (general IHC practice; HPA: tissue IHC profile). | Review blocking and antibody concentration, then compare a no-primary control with the stained section (general IHC practice). |
| Signal appears in cells HPA labels Not detected. | For example, strong bronchial respiratory epithelial staining may reflect cross-reactivity or endogenous detection activity (HPA: Not detected in bronchial respiratory epithelial cells; general IHC practice). | Inspect a no-primary control and confirm cell identity and compartment; compare a known positive population on the same run where available (general IHC practice; HPA: tissue IHC). |
| Only nuclei appear positive. | An isolated nuclear pattern does not match the supplied cytoplasmic tissue profile or UniProt localization (HPA: tissue IHC profile; UniProt Q16658: localization). | Check counterstain and chromogen appearance, then repeat interpretation against a known cytoplasmic positive control (general IHC practice; HPA: tissue IHC profile). |
| A peripheral rim is the dominant signal. | FSCN1 can associate with the cell cortex, but HPA's tissue profile emphasizes cytoplasm; its ICC-IF plasma membrane assignment is uncertain (UniProt Q16658: localization; HPA: tissue IHC and subcellular location). | Look for accompanying cytoplasmic signal and compare cell morphology and controls before calling the rim specific (general IHC practice; HPA: tissue IHC profile). |
| A whole organ is called negative despite scattered positive cells. | HPA reports expression in named cell populations, including glial cells in cerebral cortex and cells in kidney glomeruli (HPA: High in these populations). | Re-examine the relevant compartment at suitable magnification and record the positive cell population separately from surrounding tissue (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing FSCN1 staining
M02147-1 · IHCIHC analysis of Fascin using anti-Fascin antibody (M02147-1). Fascin was detected in a paraffin-embedded section of human tonsil tissue. Hea
M02147-1 · IHCIHC analysis of Fascin using anti-Fascin antibody (M02147-1). Fascin was detected in a paraffin-embedded section of mouse spleen tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000075618-FSCN1/tissue/cerebral+cortex#imid_19670643
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000075618-FSCN1/tissue/cerebellum#imid_19670630
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000075618-FSCN1/tissue/colon#imid_19670553
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000075618-FSCN1/tissue/liver#imid_19670554
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB035991; green = FSCN1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB035991; green = FSCN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for FSCN1 IHC & IF
🧪Run kidney first: cells in glomeruli should stain (HPA: High in cells in glomeruli). Use bronchus respiratory epithelial cells as the negative tissue (HPA: Not detected); on the kidney slide, cells without the glomerular staining pattern should show only background and counterstain, but the supplied HPA row does not establish them as FSCN1-negative.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Bronchus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show FSCN1 in A-431, U-251MG, U2OS, THP-1, with annotated localisation: Plasma membrane (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an irrelevant primary matched to the catalog antibody’s host species and isotype; and an FSCN1-knockout biological negative. For kidney sections developed with HRP/DAB, quench endogenous peroxidase and check for residual background (selected-SKU tissue-IHC caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state its fixative (selected-SKU tissue-IHC caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used for a paraffin section, but retrieval dependency has not been established (selected-SKU tissue-IHC caption: heat retrieval in EDTA, pH 8.0). The supplied evidence does not establish whether frozen sections or IF are easier; in kidney, check glomerular staining against residual peroxidase background (HPA: High in cells in glomeruli; selected-SKU tissue-IHC caption: HRP/DAB detection).
HPA tissue IHC evidence for FSCN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images. Paired antibodies with high similarity supports the protein expression pattern.). 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Processes in white matter | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FSCN1 IHC Tips
Troubleshoot FSCN1 staining in paraffin sections by checking retrieval, cell type, subcellular pattern, and controls before changing scoring thresholds.
Which retrieval conditions should I try first for weak FSCN1 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M02147-1). The demonstrated tonsil workflow pairs this retrieval with 1:50 primary antibody overnight at 4°C, so retain those conditions while diagnosing weak signal (datasheet M02147-1). Because the caption gives no heating duration, record the actual temperature and time, then compare retrieval times on adjacent sections with identical detection chemistry (datasheet M02147-1; general IHC practice). Include a known positive tonsil section and a no-primary control; if signal remains weak, adjust retrieval time while watching for tissue damage and rising background (datasheet M02147-1; general IHC practice).
How should I investigate inconsistent FSCN1 staining between paraffin blocks?
The selected paraffin-section example does not state a fixative, so FSCN1-specific fixation sensitivity is unknown (datasheet M02147-1). Record each block’s fixative, fixation duration, section age, and processing history before comparing staining; matched processing limits a common IHC confound (general IHC practice). Begin with the documented EDTA pH 8.0 retrieval and 1:50 primary incubation overnight at 4°C (datasheet M02147-1). If results differ, compare sections with known processing histories and assess morphology alongside staining; HPA expression patterns do not establish how fixation affects this antibody’s epitope (HPA tissue IHC; general IHC practice).
Where should FSCN1 stain, and when is nuclear signal suspicious?
Expect mainly cytoplasmic staining, potentially enriched at the cortex or cell projections: FSCN1 is annotated in cytosol, cortex, stress fibers, filopodia, and invadopodia (UniProt Q16658 subcellular). HPA tissue IHC reports cytoplasmic expression in glial cells, endothelial cells, fibroblasts, and macrophages, providing cell-level context (HPA tissue IHC). An isolated nuclear pattern is unsupported by those localisation records; check counterstained morphology and a no-primary control before scoring it (UniProt Q16658 subcellular; HPA tissue IHC; general IHC practice). Compare staining within matched cell populations using the catalog example’s 1:50 overnight incubation at 4°C as a starting condition (datasheet M02147-1; general IHC practice).
Could isoforms or modification explain patchy FSCN1 immunostaining?
The supplied UniProt record lists 0 annotated isoforms, so it does not provide an isoform-specific explanation for patchy staining (UniProt Q16658 isoforms). FSCN1 has annotated modifications, including phosphoserines at positions 38 and 39, but the supplied antibody evidence does not map its epitope or establish modification-dependent recognition (UniProt Q16658 modified residues; datasheet M02147-1). First compare retrieval, section integrity, and the expected cell populations across adjacent sections under identical staining conditions (general IHC practice; HPA tissue IHC). If an epitope effect remains a concern, seek antibody-specific epitope evidence and compare independently validated staining patterns before assigning patchiness to a modification (general IHC practice).
How can I check the FSCN1 pattern by immunofluorescence?
Treat IF as a separate validation experiment: the supplied catalog example demonstrates chromogenic IHC in a paraffin section, while HPA lists cell lines with ICC/IF images (datasheet M02147-1; HPA subcellular). Multiplex FSCN1 with a marker for the cell population under study, such as an endothelial cell marker when examining vessels, and include single-stain controls (HPA tissue IHC; general IF practice). Choose a fluorophore channel after checking tissue autofluorescence, using a longer-wavelength channel if shorter-wavelength background obscures signal (general IF practice). FSCN1 lacks a transmembrane segment and is annotated in cytosol and cell projections, so use controlled permeabilisation to access intracellular epitopes and compare it with an unpermeabilised control (UniProt Q16658 topology and subcellular; general IF practice).
How do I distinguish FSCN1 staining from diffuse DAB background?
The selected IHC example uses 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, and DAB development (datasheet M02147-1). Keep those conditions consistent while checking wash quality, primary concentration, and development time across adjacent sections (general IHC practice). Include a no-primary control and a routine peroxidase block to identify detection background from endogenous enzyme activity; these are general chromogenic IHC steps (general IHC practice). Judge remaining signal against cell morphology and the expected cytoplasmic pattern, especially in endothelial cells, fibroblasts, and macrophages that can contribute genuine FSCN1 staining (HPA tissue IHC; general IHC practice).
What is a defensible way to score FSCN1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and staining compartment before scoring, because FSCN1 is cytoplasmic and HPA reports expression in several distinct cell types (UniProt Q16658 subcellular; HPA tissue IHC). For a cell-based readout, report the percentage of positive cells and an H-score using intensity grades 0–3, yielding a range of 0–300 (general IHC practice). For sparse positive populations, report positive cells per mm² of viable, evaluable tissue and state the counting area (general IHC practice). Normalise comparisons to the same cell type or tissue compartment, section area, staining run, and scoring threshold, with positive and no-primary controls reviewed alongside samples (general IHC practice).
When should an apparent FSCN1-positive region be treated as artefact?
A credible result follows cell morphology and the chiefly cytoplasmic or projection-associated FSCN1 pattern, rather than isolated nuclear staining (UniProt Q16658 subcellular; HPA tissue IHC). Check which cells are stained: HPA reports high signal in tonsil squamous epithelial cells and cytoplasmic expression in macrophages, so staining in these populations can be genuine (HPA tissue IHC). Distrust staining confined to section edges, damaged or necrotic areas, or a no-primary control, and inspect a routine peroxidase-block control when DAB background is suspected (general IHC practice). Confirm the pattern on an adjacent section processed with the documented EDTA pH 8.0 retrieval before interpreting a discrepant region (datasheet M02147-1; general IHC practice).
Boster reagents
Best FSCN1 / Fascin IHC Antibodies
M02147-1 has IHC images of paraffin sections from human tonsil and mouse and rat spleen (catalog IHC image captions). IF/ICC is listed, but no IF image is supplied (catalog applications; IF image alts).
Real IHC data
Anti-Fascin Monoclonal Antibody
Cat # M02147-1
M02147-1 has IHC images of paraffin sections from human tonsil, mouse spleen, and rat spleen (catalog IHC image captions). It lists IHC and IF/ICC applications and human, mouse, and rat reactivity; no IF image is supplied (catalog applications; catalog reactivity; IF image alts).
Which to pick: For tissue IHC, choose M02147-1: its own captions document paraffin sections, 1:50 primary dilution, EDTA retrieval at pH 8.0, and DAB detection; the fixative is unreported (M02147-1 IHC image captions). For IF/ICC, M02147-1 is the listed option, with a catalog dilution of 1:50, although no IF figure is supplied (catalog applications; catalog IF dilution; IF image alts). For cross-species IHC, this rabbit monoclonal clone ADOA-6 lists human, mouse, and rat reactivity and has an IHC image for each species (catalog host/clone/reactivity; M02147-1 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16658 (FSCN1_HUMAN, Fascin).
- Human Protein Atlas. FSCN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. FSCN1 subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol..
- Human Protein Atlas. FSCN1 antibody validation summary (5 antibodies).
- miR-145 and miR-133a function as tumour suppressors and directly regulate FSCN1 expression in bladder cancer. British journal of cancer 2010 — PMC2833258.
- FSCN1 drives sarcoma progression and immunosuppression revealed by pan-cancer analysis and functional assays. Discover oncology 2026 — PMC13243165.
- Fascin-1 expression in non-small cell lung cancer and its diagnostic implications. Discover oncology 2026 — PMC13461984.
- FSCN1 is an effective marker of poor prognosis and a potential therapeutic target in human tongue squamous cell carcinoma. Cell death & disease 2019 — PMC6494834.
- PubMed PMID:8068206 — UniProt-cited evidence.
- PubMed PMID:7933116 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for FSCN1 (Fascin). Expected localisation Cytoplasmic staining in tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02147-1). Validated IHC antibody: M02147-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Bronchus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet M02147-1); 2. Fixation: Image fixative and duration unreported (datasheet M02147-1); 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 M02147-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02147-1); 8. Primary antibody: Rabbit monoclonal (clone ADOA-6) anti-FSCN1, 1:50 (datasheet M02147-1); 9. Primary incubation: Overnight at 4 °C (datasheet M02147-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Macrophage and endothelial staining can complicate cell scoring (HPA tissue IHC); Isoform / epitope: No isoforms annotated; Fascin chain spans residues 2–493 (UniProt); Regulation: Low tissue specificity; staining varies by cell type (HPA tissue IHC). Evidence: UniProt Q16658, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol., Datasheet M02147-1, PMC2833258, PMC13243165. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/FSCN1/immunohistochemistry-ihc-guide.