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FUT2 / Galactoside alpha-(1,2)-fucosyltransferase 2 · IHC design guide
Design Immunohistochemistry for FUT2
Plan FUT2 staining in paraffin sections with the catalog antibody’s 1:100–1:300 IHC dilution range (datasheet A02679). Expect cytoplasmic tissue staining (HPA tissue IHC), but interpret intensity cautiously because staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
FUT2 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 | Observed cytoplasmic staining (HPA tissue IHC); expected Golgi membrane location (UniProt) | |
| Staining pattern | Cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A02679) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Vagina |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent between sections (standard IHC practice; not target-specific) | |
| Caveat | Low consistency between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Tissue-enhanced RNA in intestine (HPA tissue RNA) | |
| Isoform / epitope | No isoforms annotated; cytosolic versus lumenal epitope matters (UniProt) |
Section 1
Recommended FUT2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A02679) is accompanied by published FUT2 IHC methods for lung adenocarcinoma (PMC5722559) and colorectal cancer (PMC9911090).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02679) |
| Fixation | Image fixative and duration unreported (datasheet A02679); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A02679); 20 min, 95–100 °C (standard) |
| 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-FUT2, 1:100-1:300 (datasheet A02679) |
| 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 | FUT2-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 for the catalog antibody (datasheet A02679); the published methods use sodium citrate retrieval (PMC5722559; PMC9911090).
Section 2
What Is the Expected FUT2 Staining Pattern?
FUT2 is a Golgi stack membrane enzyme with a cytoplasmic N terminus and a lumenal catalytic region (UniProt Q10981 topology). In paraffin IHC, assess cytoplasmic staining in glandular or respiratory epithelial cells; appendix glandular and bronchial epithelial cells show medium staining (HPA tissue IHC). Treat the pattern as provisional: HPA rates tissue staining reliability Uncertain because staining and RNA expression have low consistency (HPA tissue IHC).
| Localized cytoplasmic staining in appendix glandular cells or bronchial epithelium. | This agrees with medium staining reported in those cells (HPA tissue IHC). A compact perinuclear component is compatible with Golgi localization (UniProt Q10981 subcellular location). Compare the signal with adjacent tissue and a no-primary control before calling cells positive (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear pattern is unsupported by the Golgi membrane annotation (UniProt Q10981 subcellular location). Check the no-primary control and review whether chromogen precipitate or counterstain is being read as signal (general IHC practice). Membrane staining alone needs separate assessment because HPA reports an uncertain plasma membrane ICC-IF location (HPA ICC-IF). |
| Strong staining in vaginal squamous epithelium, especially if the expected control is weak. | HPA reports FUT2 as not detected in vaginal squamous cells (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity as possible IHC explanations (general IHC practice); this comparison cannot prove either mechanism because HPA rates tissue staining Uncertain (HPA tissue IHC). |
| Even haze across tissue, stroma, and empty spaces. | A slide-wide haze lacks the cell-specific pattern needed to interpret FUT2 staining (HPA tissue IHC; general IHC practice). Inspect no-primary and reagent controls, then optimize blocking, washes, antibody concentration, and detection conditions one at a time (general IHC practice). |
| No visible signal in appendix glandular cells or bronchial epithelium. | These are reported medium-staining populations, making them useful comparison tissues, although the HPA tissue result is Uncertain (HPA tissue IHC). Check tissue preservation, retrieval conditions, reagent activity, and a run-level positive control before treating the absence of stain as biological (general IHC practice). |
💡Expected FUT2 appearanceCall a result provisionally positive when glandular or respiratory epithelial cells show distinct cytoplasmic, potentially perinuclear staining around HPA's medium level (HPA tissue IHC; UniProt Q10981 subcellular location); nuclear signal or uniform slide-wide haze is a warning sign (UniProt Q10981 subcellular location; general IHC practice).
| Compartment and epitope orientation | FUT2 spans the membrane at residues 15–28; residues 1–14 are cytoplasmic and 29–343 are lumenal (UniProt Q10981 topology). The supplied record gives no antibody epitope, so orientation cannot establish a required retrieval method or staining intensity. |
| Tissue and antibody evidence | HPA calls tissue staining Uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC). Its listed antibody HPA014402 has Uncertain IHC validation (HPA antibody validation). Use reported tissue levels as comparisons, not definitive specificity proof. |
| IF/ICC Q&A: Should membrane signal match IHC? | HPA places FUT2 mainly at the plasma membrane, with additional cytosol, in ICC-IF; both locations are uncertain (HPA ICC-IF). Tissue IHC instead reports cytoplasmic expression (HPA tissue IHC), while UniProt places FUT2 at Golgi membranes (UniProt Q10981 subcellular location). |
| Processing and glycosylation | UniProt lists no signal peptide or propeptide and annotates glycosylation at residues 188, 282, and 308 (UniProt Q10981 processing and glycosylation). Neither annotation establishes a fixation effect, retrieval requirement, or cause of a negative IHC result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected epithelial control is blank. | The run may have a tissue, retrieval, antibody, or detection problem (general IHC practice); appendix and bronchus are only provisional medium-positive references (HPA tissue IHC). | Confirm tissue identity and a working run-level control; review retrieval conditions, primary antibody use, and detection reagents in sequence (general IHC practice). |
| Nuclear staining dominates. | Nuclear localization is unsupported for FUT2 (UniProt Q10981 subcellular location); counterstain or deposited chromogen may be misread as target signal (general IHC practice). | Compare the no-primary section and inspect signal separately from counterstain; reassess the result against the expected cytoplasmic distribution (HPA tissue IHC; general IHC practice). |
| Vaginal squamous cells appear strongly positive. | This conflicts with HPA's not-detected observation (HPA tissue IHC). Cross-reactivity or endogenous detection activity are possibilities, not established causes (general IHC practice). | Check a no-primary control and the detection system's endogenous-activity control; compare with a reported medium-positive tissue (HPA tissue IHC; general IHC practice). |
| Diffuse staining obscures cell boundaries. | Excess primary or detection reagent, insufficient blocking, or inadequate washing can raise background (general IHC practice). | Compare reagent controls, then adjust blocking, primary concentration, detection exposure, and washes separately while retaining the same tissue control (general IHC practice). |
| Only surface membrane outlines are visible. | HPA's plasma membrane ICC-IF assignment is uncertain and does not establish the expected paraffin IHC appearance (HPA ICC-IF; HPA tissue IHC). | Review cytoplasmic signal in the same cells and compare controls; record membrane-only staining as unresolved unless independently supported (UniProt Q10981 subcellular location; general IHC practice). |
| Staining varies sharply between sections. | Section condition or run handling may differ (general IHC practice); HPA's Uncertain tissue reliability also limits confidence in a biological explanation (HPA tissue IHC). | Compare matched sections with the same control tissue and document retrieval, antibody application, detection time, and counterstain for each run (general IHC practice). |
Experiment evidence
IHC & IF images showing FUT2 staining
A02679 · IHCImmunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176920-FUT2/tissue/cerebral+cortex#imid_3796800
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000176920-FUT2/tissue/duodenum#imid_3796725
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176920-FUT2/tissue/colon#imid_3796734
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176920-FUT2/tissue/liver#imid_3796735
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA014402; green = FUT2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA014402; green = FUT2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for FUT2 IHC & IF
🧪Run appendix first: its glandular cells should stain (HPA: Medium in appendix glandular cells). Run vagina as the negative tissue (HPA: Not detected in vaginal squamous epithelial cells); on the appendix slide, cells outside the stained glands should lack specific signal.
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Vagina (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show FUT2 in A-431, CACO-2, U2OS, with annotated localisation: Plasma membrane (uncertain) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and primary-host-matched isotype controls; for a polyclonal primary, use nonimmune serum from the same host species. Use matched FUT2-knockout tissue as a biological negative where available, and check the appendix slide for endogenous peroxidase signal and background in gland lumens (HPA: Medium in appendix glandular cells).
⚠️Feasibility: A FUT2-specific fixation window or fixation effect is unreported, and the selected A02679 paraffin-section caption does not state a fixative (A02679 tissue-IHC caption). The caption uses Tris-EDTA at pH 9.0 for retrieval in liver cancer, but retrieval dependence in appendix is unreported (A02679 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; in appendix, distinguish glandular-cell staining from luminal background (HPA: Medium in appendix glandular cells).
HPA tissue IHC evidence for FUT2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FUT2 IHC Tips
Use the paraffin-section IHC conditions reported for A02679 as a starting point, and assess staining against FUT2 localisation and tissue context.
What retrieval should I use if FUT2 staining is weak in paraffin sections?
Use heat-mediated Tris-EDTA at pH 9.0 for paraffin sections (datasheet A02679). The selected tissue-IHC image used that retrieval with the primary antibody at 1:200, incubated at 4°C overnight (caption A02679). If staining is weak, check heating consistency, section adhesion and whether the positive control stains before changing retrieval conditions (standard IHC practice). If you test a different buffer or pH, compare adjacent sections with the documented condition and the same detection settings; the caption does not establish that an alternative improves FUT2 detection (caption A02679; standard IHC practice).
How should I troubleshoot fixation-related loss of FUT2 signal?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A02679). Record the specimen’s actual fixative and fixation duration, then compare similarly processed sections before attributing a weak stain to fixation (standard IHC practice). Keep section thickness, heat retrieval in Tris-EDTA at pH 9.0 and antibody incubation consistent across that comparison (datasheet A02679; standard IHC practice). Include a processing-matched positive control and inspect its staining alongside the test section; tissue patterns and FUT2 topology do not establish how this antibody responds to any particular fixative (HPA tissue IHC; UniProt Q10981 topology; standard IHC practice).
Should FUT2 appear at the cell surface or around the Golgi in IHC?
Evaluate a compact intracellular staining pattern in the context of FUT2’s Golgi stack membrane annotation, including its reported trans-cisternal membrane-bound form (UniProt Q10981 localisation). Its 15–28 transmembrane segment anchors a short cytoplasmic region and a larger lumenal region, so antibody epitope accessibility may affect the observed pattern (UniProt Q10981 topology; standard IHC practice). HPA reports cytoplasmic tissue staining, while its plasma-membrane and cytosol IF assignments are uncertain (HPA tissue IHC; HPA subcellular). Review staining within identifiable cells, morphology and matched controls before calling a diffuse or membrane-dominant chromogenic pattern specific; compartment appearance alone cannot resolve the differing annotations (standard IHC practice; HPA subcellular).
Could FUT2 processing or epitope position explain discrepant IHC staining?
The supplied UniProt record annotates 0 isoforms and a 1–343 chain, so an isoform-specific explanation has no support here (UniProt Q10981 isoforms and processing). FUT2 has a cytoplasmic segment at 1–14, a transmembrane segment at 15–28 and a lumenal region at 29–343 (UniProt Q10981 topology). Glycosylation is annotated at 188, 282 and 308, but the catalog antibody’s binding epitope is unspecified (UniProt Q10981 glycosylation; supplied catalog evidence). Ask for epitope information before assigning a loss of staining to masking or glycosylation, and compare retrieval conditions on matched sections with consistent controls (standard IHC practice).
How can IF help assess a questionable FUT2 IHC pattern?
For a separate IF assessment, multiplex FUT2 with an epithelial marker in appendix glandular cells, which have medium tissue-IHC staining in HPA (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, favoring a far-red channel when tissue autofluorescence obscures shorter wavelengths, and include single-stain and no-primary controls (standard IF practice). If the antibody recognizes the lumenal 29–343 region of Golgi-bound FUT2, permeabilisation is needed to access that intracellular epitope; verify the antibody’s actual epitope first (UniProt Q10981 topology; standard IF practice). Judge colocalisation against cell morphology and compartment pattern, while treating HPA’s plasma-membrane and cytosol IF assignments as uncertain (HPA subcellular; standard IF practice).
How do I reduce widespread brown signal without losing FUT2 staining?
Start by checking a no-primary control and the tissue’s endogenous peroxidase activity before interpreting DAB color as FUT2 (standard chromogenic IHC practice). Use an appropriate peroxidase block, protein block and thorough washes, then compare sections processed with the documented 1:200 primary dilution and 1:200 secondary dilution (caption A02679; standard IHC practice). The selected image used a 45-minute room-temperature secondary incubation; it does not show that stronger detection is more specific (caption A02679). If background persists, adjust one detection variable at a time while preserving a processing-matched positive control, and exclude edge deposits or damaged areas from interpretation (standard IHC practice).
How should I score FUT2 when staining varies across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, and keep imaging, counterstain and threshold settings consistent across sections (standard IHC quantification practice). For glandular tissue, report the percentage of positive glandular cells and an H-score from 0–300 based on staining intensity; document the intensity bins used (standard IHC quantification practice). Normalize positive counts to the number of evaluable cells in that compartment, or report positive-cell density per mm² of viable, annotated tissue (standard IHC quantification practice). Exclude folds, necrosis and section edges, and report replicate sections or specimens separately because HPA rates its FUT2 tissue-IHC reliability uncertain (standard IHC practice; HPA tissue IHC).
What distinguishes convincing FUT2 positivity from staining artefact?
A convincing result should stain intact cells reproducibly, agree with a processing-matched control and be assessed against FUT2’s Golgi membrane annotation (UniProt Q10981 localisation; standard IHC practice). Examine the cell identity: HPA reports medium staining in appendix glandular cells, but rates overall tissue-IHC reliability uncertain because staining and RNA data have low consistency (HPA tissue IHC). Treat signal confined to cut edges, necrotic regions or endogenous-peroxidase-positive areas as suspect, and compare it with no-primary and blocked controls (standard chromogenic IHC practice). A membrane-dominant pattern needs further validation because HPA’s plasma-membrane IF assignment is uncertain; the paraffin liver-cancer caption documents staining conditions, not specificity by itself (HPA subcellular; caption A02679).
Boster reagents
Best FUT2 / Galactoside alpha-(1,2)-fucosyltransferase 2 IHC Antibodies
A02679 has real IHC data from paraffin-embedded human liver cancer (IHC image caption). IF is listed, with human, mouse, and rat reactivity (catalog: applications/reactivity).
Real IHC data
Anti-FUT2 Antibody
Cat # A02679
A02679 shows IHC staining of paraffin-embedded human liver cancer using Tris-EDTA retrieval at pH 9.0 and a 1:200 primary dilution (IHC image caption). The catalog lists IHC and IF applications and human, mouse, and rat reactivity, but provides no IF image (catalog: applications/reactivity/IF image alts).
Which to pick: Choose A02679 for paraffin-section tissue IHC based on its human liver cancer image; the fixative is unreported (A02679 IHC image caption). For IF, A02679 is listed at 1:50; ICC validation and IF images are unreported (catalog: applications/IF dilution/IF image alts). For mouse or rat samples, A02679 lists reactivity with both species, although its IHC image shows human tissue only; the antibody is rabbit-hosted and its clonality is unreported (catalog: reactivity/host/clone; A02679 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q10981 (FUT2_HUMAN, Galactoside alpha-(1,2)-fucosyltransferase 2).
- Human Protein Atlas. FUT2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. FUT2 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the cytosol..
- Human Protein Atlas. FUT2 antibody validation summary (1 antibodies).
- Clinical significance and biological function of fucosyltransferase 2 in lung adenocarcinoma. Oncotarget 2017 — PMC5722559.
- FUT2 enhances anti-tumor immunity in pancreatic cancer radiotherapy by driving FBXO2-mediated degradation of NR2F2. Cell death & disease 2025 — PMC12848027.
- A Glycosyltransferase-Related Signature for Predicting Overall Survival in Head and Neck Squamous Cell Carcinoma. Frontiers in genetics 2022 — PMC9311713.
- FUT2 promotes the tumorigenicity and metastasis of colorectal cancer cells via the Wnt/β‑catenin pathway. International journal of oncology 2023 — PMC9911090.
- PubMed PMID:7876235 — UniProt-cited evidence.
- PubMed PMID:9219535 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for FUT2 (Galactoside alpha-(1,2)-fucosyltransferase 2). Expected localisation Observed cytoplasmic staining (HPA tissue IHC); expected Golgi membrane location (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A02679). Validated IHC antibody: A02679 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Vagina. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A02679); 2. Fixation: Image fixative and duration unreported (datasheet A02679); 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: Tris-EDTA pH 9.0 (datasheet A02679); 20 min, 95–100 °C (standard); 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-FUT2, 1:100-1:300 (datasheet A02679); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent between sections (standard IHC practice; not target-specific); Caveat: Low consistency between staining and RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms annotated; cytosolic versus lumenal epitope matters (UniProt); Regulation: Tissue-enhanced RNA in intestine (HPA tissue RNA). Evidence: UniProt Q10981, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A02679, PMC5722559, PMC9911090. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/FUT2/immunohistochemistry-ihc-guide.