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FUT6 / 4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase FUT6 · IHC design guide
Design Immunohistochemistry for FUT6
Plan FUT6 paraffin IHC around granular cytoplasmic staining in gastrointestinal tissue (HPA tissue IHC). Start the IHC-validated antibody at 1:100–1:200 (datasheet: A06862), and account for a secreted variant when interpreting tissue staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
FUT6 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 | Granular cytoplasm in tissue (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm in enterocytes and glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | A secreted variant may shift tissue staining (HPA tissue IHC) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended FUT6 IHC & IF Protocols
The catalog antibody IHC-P protocol is followed by published FUT6 staining protocols for prostate biopsies and mouse tumor sections (PMC3699712; PMC3847326; PMC7811455).
| Sample | Paraffin-embedded human liver tissue; fixative not specified (datasheet A06862) |
| Fixation | Image fixative and duration unreported (datasheet A06862); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-FUT6, 1:100-1:200 (datasheet A06862) |
| 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 | FUT6-positive staining in enterocytes - Microvilli of duodenum (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in gastrointestinal tract, gallbladder, seminal vesicle and cervix. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); optimize against the published conditions below.
Section 2
What Is the Expected FUT6 Staining Pattern?
FUT6 is a Golgi stack membrane protein with a lumenal catalytic region and a reported secreted form (UniProt P51993 topology; UniProt P51993 subcellular location). In paraffin section IHC, expect granular cytoplasmic staining in gastrointestinal cells and strong staining at duodenal enterocyte microvilli (HPA tissue IHC). HPA rates its tissue IHC profile Approved, with medium consistency against RNA and a caution that antibodies may detect proteins from more than one gene (HPA tissue IHC).
| Granular cytoplasmic staining in gastrointestinal cells, with strong duodenal enterocyte microvilli staining (HPA tissue IHC). | This matches HPA’s tissue profile. Supported Golgi localization gives the cytoplasmic granules a plausible compartment, while the observed microvilli pattern should be recorded separately from Golgi staining (HPA tissue IHC; HPA ICC-IF; UniProt P51993 subcellular location). |
| Predominantly nuclear staining, without the expected gastrointestinal cytoplasmic pattern. | A nuclear-dominant pattern is unsupported by the Golgi localization records (UniProt P51993 subcellular location; HPA ICC-IF). Check the no-primary control and staining specificity before scoring it as FUT6; the observation alone does not identify the artefact (general IHC practice). |
| Prominent staining in a cell population HPA reports as not detected, such as adipocytes in adipose tissue. | Treat this as discordant with the reported IHC pattern, not automatic proof of cross-reactivity (HPA tissue IHC). Compare morphology and controls; off-target binding or endogenous detection activity are possible explanations in chromogenic IHC (general IHC practice). |
| Diffuse color covers cells and surrounding tissue, obscuring the granular pattern. | That appearance is difficult to score against HPA’s granular cytoplasmic profile (HPA tissue IHC). Uneven blocking, residual detection reagent or excessive chromogen development can create background; inspect controls and the detection workflow (general IHC practice). |
| No signal in duodenal enterocytes on a run expected to show FUT6. | HPA reports high staining at duodenal enterocyte microvilli, so a blank result warrants a run check (HPA tissue IHC). Confirm tissue preservation and morphology, then review retrieval, antibody dilution and detection controls; none has a supplied FUT6-specific setting (general IHC practice). |
💡Expected FUT6 appearanceCall a positive result when gastrointestinal cells show interpretable granular cytoplasmic staining and duodenal enterocytes show strong microvilli staining; nuclear-only color or uniform diffuse deposit is discordant (HPA tissue IHC; HPA ICC-IF; general IHC practice).
| Cell and compartment choice | Duodenal enterocyte microvilli are High; gallbladder glandular cells and colonic endocrine cells are Medium (HPA tissue IHC). Golgi localization is supported independently (HPA ICC-IF; UniProt P51993). |
| Antibody interpretation | HPA calls the tissue IHC profile Approved, with medium RNA consistency and a multiple-gene detection caution; its listed IHC antibody is Approved, not Enhanced (HPA tissue IHC; HPA antibodies). |
| Membrane topology and epitope | Residues 15–34 span the membrane; residues 35–359 are lumenal (UniProt P51993 topology). Epitope location is not supplied, so topology alone cannot specify retrieval or staining intensity. |
| Secreted form | UniProt reports both membrane-bound Golgi FUT6 and a secreted form (UniProt P51993 subcellular location). Extracellular color needs cautious interpretation because the tissue IHC profile does not establish its source (HPA tissue IHC). |
| Variants and glycosylation | Two isoforms and glycosylation sites at residues 46, 91, 153 and 184 are annotated (UniProt P51993). Their effects on this antibody’s paraffin section staining are unreported. |
| RNA versus protein | Esophagus is RNA-enhanced, yet its squamous epithelial cells are reported Not detected by IHC (HPA tissue IHC). Choose and score controls using the protein staining record, with HPA’s RNA consistency caveat (HPA tissue IHC). |
| IF/ICC: what pattern is supported? | Golgi localization is supported in ICC-IF images from A-431, Hep-G2 and U2OS (HPA ICC-IF). This is a localization reference, not an IHC-P protocol or evidence of identical intensity in sections. |
| Situation | Likely cause | Next action |
|---|---|---|
| Duodenum is blank. | A run or reagent problem is possible because duodenal enterocyte microvilli are reported High (HPA tissue IHC). | Inspect morphology and a run control; review retrieval, antibody dilution and detection in the catalog antibody’s IHC-P protocol (general IHC practice). |
| Granular cytoplasmic signal is weak but tissue morphology is intact. | The sampled cells may differ from the stronger reference: HPA reports Medium staining in several listed gastrointestinal cell populations and High at duodenal microvilli (HPA tissue IHC). | Confirm the cell type and compare with a duodenum positive control before changing the IHC workflow (HPA tissue IHC; general IHC practice). |
| Nuclei dominate the stain. | That distribution conflicts with supported Golgi localization; its precise cause cannot be assigned from appearance alone (HPA ICC-IF; UniProt P51993). | Check no-primary and detection controls, then verify whether cytoplasmic granules are present on a positive tissue (general IHC practice; HPA tissue IHC). |
| A nominal negative tissue stains strongly. | HPA warns of detection from more than one gene; endogenous chromogenic activity or nonspecific binding can also give color (HPA tissue IHC; general IHC practice). | Compare the same cell type with HPA’s IHC record and review no-primary, blocking and detection controls before calling FUT6 positive (HPA tissue IHC; general IHC practice). |
| Color spreads uniformly across the section. | Diffuse deposit can hide the reported granular pattern; reagent background or excessive development is possible (HPA tissue IHC; general IHC practice). | Review blocking, washes and chromogen development against run controls, then score only localized cellular signal (general IHC practice). |
| RNA suggests a positive tissue, but IHC is negative. | RNA and protein patterns need not agree; HPA specifically reports RNA-enhanced esophagus with squamous epithelial IHC Not detected (HPA tissue IHC). | Use HPA’s cell-level protein pattern for the IHC comparison, and document the RNA/IHC discrepancy rather than treating RNA as a positive stain control (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing FUT6 staining
A06862 · IHCImmunohistochemistry of paraffin-embedded human liver using FUT6 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10
A06862 · IHCImmunohistochemistry of paraffin-embedded human stomach using FUT6 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 1
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156413-FUT6/tissue/cerebral+cortex#imid_14002105
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156413-FUT6/tissue/colon#imid_13999954
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156413-FUT6/tissue/liver#imid_13999955
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156413-FUT6/tissue/kidney#imid_13999956
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043707; green = FUT6, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043707; green = FUT6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for FUT6 IHC & IF
🧪Run duodenum first: enterocyte microvilli should stain (HPA: High in duodenal enterocyte microvilli). Use adipose tissue as the negative tissue, with adipocytes expected at background (HPA: Not detected in adipocytes); on the duodenal slide, use neighboring non-enterocyte profiles as an internal background comparison, without assuming every such cell is FUT6-negative.
Positive control tissue: Duodenum (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show FUT6 in A-431, Hep-G2, U2OS, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-matched, clonality-matched isotype controls; use FUT6 knockout material as a biological negative where available (standard IHC control practice). For chromogenic staining, quench endogenous peroxidase and check for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected paraffin-section caption does not state a fixative (A06862 caption). Its reported starting condition is microwave antigen retrieval in 10 mM PBS, pH 7.2, followed by antibody at 1:100; retrieval dependency beyond that example is unreported (A06862 caption). The evidence does not establish whether frozen sections or IF are easier; in duodenum, distinguish microvillar staining from diffuse luminal signal when scoring (HPA: High in duodenal enterocyte microvilli).
HPA tissue IHC evidence for FUT6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Duodenum | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Colon | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FUT6 IHC Tips
Troubleshoot FUT6 staining by checking retrieval, compartment, cell type and controls together (UniProt P51993; HPA tissue IHC).
How should I adjust retrieval when FUT6 staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval rule). If staining remains weak, test microwave retrieval in 10 mM PBS, pH 7.2 on a matched section; that condition accompanies the paraffin embedded human liver image for catalog antibody A06862 (A06862 tissue IHC caption). Keep the antibody dilution, detection system and development time constant while comparing retrieval conditions (standard IHC practice). Include a no primary control and examine tissue morphology, since stronger colour alone does not establish specific FUT6 detection (standard IHC practice).
Could fixation explain weak or uneven FUT6 staining?
Target specific fixation sensitivity is unknown: the selected paraffin section caption does not state a fixative (A06862 tissue IHC caption). Record the fixative and fixation duration for each specimen, then compare similarly processed sections before changing retrieval or antibody concentration (standard IHC practice). For uneven staining, inspect folds, section thickness and tissue edges, and assess whether the pattern follows processing rather than cell boundaries (standard IHC practice). Use a documented positive control alongside each run; duodenal enterocytes provide a reported high staining reference, although the assay's specificity still requires controls (HPA tissue IHC: High in duodenal enterocytes; HPA tissue IHC reliability: Approved).
Which staining compartment should count as plausible FUT6 signal?
Expect a predominantly granular cytoplasmic pattern compatible with Golgi localisation, while allowing for the reported secreted form (HPA tissue IHC: granular cytoplasmic expression; HPA subcellular: Golgi apparatus supported; UniProt P51993 localisation). Membrane bound FUT6 occupies trans Golgi cisternae, with residues 1–14 cytoplasmic and 35–359 lumenal (UniProt P51993 topology). Score a diffuse nuclear signal cautiously and check its presence in the no primary control before calling cells positive (UniProt P51993 localisation; standard IHC practice). Compare the compartment and cell type across serial sections rather than treating any brown cytoplasm as proof of FUT6 (standard IHC practice).
How do isoforms and epitope position affect my IHC interpretation?
FUT6 has 2 annotated isoforms, but the supplied antibody information does not identify its epitope or establish isoform coverage (UniProt P51993 isoforms; A06862 tissue IHC caption). Check the antibody's epitope documentation before claiming that staining measures both isoforms (standard IHC practice). FUT6 spans the membrane at residues 15–34, leaving a short cytoplasmic segment and a long lumenal region with glycosylation sites at 46, 91, 153 and 184 (UniProt P51993 topology and glycosylation). If two antibodies give different patterns, compare their documented epitopes and matched controls before attributing the difference to isoform expression or glycosylation (standard IHC practice).
How can IF help check a questionable chromogenic FUT6 pattern?
Use IF as an independent spatial check and multiplex FUT6 with a marker for the expected cell type, such as an enterocyte marker when examining duodenum (HPA tissue IHC: High in duodenal enterocytes; standard IF practice). Choose fluorophores after measuring tissue autofluorescence and include single label and no primary controls (standard IF practice). Permeabilise sufficiently for the antibody's documented epitope: residues 1–14 face the cytoplasm, whereas residues 35–359 face the Golgi lumen; the supplied caption does not locate this antibody's epitope (UniProt P51993 topology; A06862 tissue IHC caption). Assess Golgi pattern agreement without treating IF intensity as interchangeable with chromogenic IHC intensity (HPA subcellular: Golgi apparatus supported; standard IF practice).
What should I check when FUT6 staining is widespread or muddy?
Run a no primary control to assess detection reagent signal, and inspect whether brown deposits concentrate at edges, folds or damaged areas (standard IHC practice). Block endogenous peroxidase before chromogenic detection and titrate antibody concentration or development time against a documented positive section (standard IHC practice). The selected image used antibody A06862 at 1:100 in paraffin embedded human liver, which is an image condition rather than a validated optimum for every specimen (A06862 tissue IHC caption). Be especially cautious with broad staining because the HPA tissue assay warns that its antibody targets protein from more than one gene (HPA tissue IHC reliability description).
How should I quantify FUT6 across sections with different cell compositions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the relevant cell population and staining compartment before scoring; HPA reports granular cytoplasmic expression and cell type dependent staining (HPA tissue IHC profile and positive cells). For chromogenic IHC, report the percentage of positive target cells and an H score using intensity categories 0–3, or positive cell density per mm² when counts are more useful (standard IHC practice). Normalise cell counts to the number of evaluable cells of that type or to analysed tissue area, and exclude folds and necrosis consistently (standard IHC practice). Keep retrieval, detection, imaging and scoring rules identical across comparisons (standard IHC practice).
When is a FUT6 positive call convincing rather than an artefact?
A convincing call combines the expected cell type with granular cytoplasmic staining compatible with Golgi localisation and a clean no primary control (HPA tissue IHC profile; HPA subcellular: Golgi apparatus supported; standard IHC practice). Duodenal enterocytes offer a reported high staining reference, while HPA reports no detection in liver cholangiocytes; these observations describe different cells from the human liver image and should not be conflated (HPA tissue IHC; A06862 tissue IHC caption). Challenge isolated nuclear staining, edge accentuation and necrotic deposits, and check endogenous peroxidase background (UniProt P51993 localisation; standard IHC practice). Interpret discordant tissue patterns cautiously because a secreted variant can complicate RNA and protein comparisons, and the HPA tissue assay has a multiple gene targeting caveat (HPA tissue IHC reliability description).
Boster reagents
Best FUT6 / 4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase FUT6 IHC Antibodies
A06862 has IHC images of human paraffin-embedded liver and stomach (catalog image captions); no IF image is supplied (catalog IF images: none).
Real IHC data
Anti-FUT6 Antibody
Cat # A06862
A06862 will render with its human liver paraffin-section IHC figure at 1:100 (catalog liver image caption). Its application list includes IHC and WB, and a separate caption reports human stomach paraffin-section IHC at 1:100 (catalog applications; catalog stomach image caption).
Which to pick: Choose rabbit antibody A06862 for human paraffin-section IHC; its captions report microwave antigen retrieval in 10 mM PBS, pH 7.2, and the catalog IHC range is 1:100–1:200 (catalog host; catalog liver and stomach image captions; datasheet: IHC dilution). The captions do not report the fixative (catalog liver and stomach image captions). No IF/ICC application or nonhuman reactivity is listed for A06862, so the payload does not support an IF/ICC or cross-species pick (catalog applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51993 (FUT6_HUMAN, 4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase FUT6).
- Human Protein Atlas. FUT6 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. FUT6 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. FUT6 antibody validation summary (2 antibodies).
- Comparing the Immunoexpression of FUT3 and FUT6 between Prostatic Adenocarcinoma and Benign Prostatic Hyperplasia. Acta histochemica et cytochemica 2013 — PMC3699712.
- FUT family mediates the multidrug resistance of human hepatocellular carcinoma via the PI3K/Akt signaling pathway. Cell death & disease 2013 — PMC3847326.
- CircSND1 Regulated by TNF-α Promotes the Migration and Invasion of Cervical Cancer Cells. Cancer management and research 2021 — PMC7811455.
- Fucosyltransferases Regulated by Fusobacterium Nucleatum and Act as Novel Biomarkers in Colon Adenocarcinoma. Journal of inflammation research 2023 — PMC9960735.
- PubMed PMID:1520296 — UniProt-cited evidence.
- PubMed PMID:1339443 — UniProt-cited evidence.
- PubMed PMID:7650030 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for FUT6 (4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase FUT6). Expected localisation Granular cytoplasm in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06862 at 1:100-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Enterocytes - Microvilli); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver tissue; fixative not specified (datasheet A06862); 2. Fixation: Image fixative and duration unreported (datasheet A06862); 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 citrate buffer, pH 6.0 , 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-FUT6, 1:100-1:200 (datasheet A06862); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: A secreted variant may shift tissue staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; verify antibody epitope coverage (UniProt); Regulation: Regulation not specified (UniProt). Evidence: UniProt P51993, HPA tissue IHC · Approved, HPA ICC-IF · Golgi apparatus., Datasheet A06862, PMC3699712, PMC3847326. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/FUT6/immunohistochemistry-ihc-guide.