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ST6GAL1 / Beta-galactoside alpha-2,6-sialyltransferase 1 · IHC design guide
Design Immunohistochemistry for ST6GAL1
Plan paraffin-section ST6GAL1 IHC with hepatocytes and tonsillar germinal-center cells as high-staining references (HPA tissue IHC). This guide covers the cytoplasmic Golgi pattern (HPA tissue IHC), a catalog antibody dilution of 1:100–1:300 (datasheet A30620), and interpretation of a potentially secreted form (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ST6GAL1 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 Golgi pattern (HPA tissue IHC) | |
| Staining pattern | Hepatocyte and germinal-center cell Golgi staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A30620) | |
| Positive control | Liver+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30620) | |
| Caveat | A secreted variant may alter tissue staining (HPA tissue IHC) | |
| Regulation | Liver and lymphoid tissue group enrichment (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; cytoplasmic versus luminal epitope coverage is unknown (UniProt) |
Section 1
Recommended ST6GAL1 IHC & IF Protocols
The catalog antibody’s paraffin-section protocol uses EDTA pH 8.0 retrieval (datasheet A30620). Published chromogenic protocols describe TNBC cryosections and lymphoma cytospins (PMC9663294; PMC4027875).
| Sample | Paraffin-embedded Human stomach tissue; fixative not specified (datasheet A30620) |
| Fixation | Image fixative and duration unreported (datasheet A30620); 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 A30620); 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-ST6GAL1, 1:100 - 1:300 (datasheet A30620) |
| 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 | ST6GAL1-positive staining in hepatocytes of liver (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with a Golgi pattern in several tissues, including liver and lymphoid tissues. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with heat-mediated EDTA pH 8.0 retrieval (datasheet A30620). Neither published cryosection nor cytospin method specifies retrieval (PMC9663294; PMC4027875).
Section 2
What Is the Expected ST6GAL1 Staining Pattern?
ST6GAL1 is a Golgi stack membrane protein with a lumenal catalytic region and a reported secreted form (UniProt P15907 topology and location). In IHC, expect cytoplasmic staining with a Golgi pattern, especially in hepatocytes and tonsillar germinal center cells (HPA tissue IHC: High in both). HPA rates the tissue evidence Enhanced but notes only medium consistency with RNA expression and a possible mismatch between RNA and protein location (HPA tissue IHC).
| Hepatocytes or tonsillar germinal center cells show concentrated cytoplasmic staining near the nucleus. | This fits the Golgi pattern reported across tissues and the High staining recorded in these two cell populations (HPA tissue IHC). Judge the distribution within the expected cells as well as signal strength; a dark deposit alone does not establish the expected pattern (general IHC practice). |
| The dominant deposit is nuclear, or it sharply outlines cell surfaces without a Golgi-like cytoplasmic component. | That distribution does not match the reported Golgi stack membrane location or HPA tissue pattern (UniProt P15907 location; HPA tissue IHC). Treat it as suspect and compare it with a known-positive section and appropriate detection controls before assigning it to ST6GAL1 (general IHC practice). |
| Strong staining appears in cells recorded as Not detected, such as adipocytes or cardiomyocytes. | That conflicts with the supplied tissue observations (HPA tissue IHC: Not detected in adipocytes and cardiomyocytes). Cross-reactivity or endogenous chromogenic activity is possible; a no-primary control can help distinguish detection-system signal from primary-antibody-associated staining (general IHC practice). |
| Color is spread uniformly across cells or tissue, obscuring a discrete Golgi-like pattern. | Diffuse background limits interpretation even where ST6GAL1 is expected (HPA tissue IHC: cytoplasmic Golgi pattern). Review the no-primary control, blocking, washes and chromogen development before scoring positive cells (general IHC practice); diffuse color by itself is not evidence of a secreted form. |
| No convincing stain appears in hepatocytes or tonsillar germinal center cells. | These are useful positive comparators because HPA records High staining in both (HPA tissue IHC). A blank result there raises a run-level or antibody-use question before a negative result elsewhere is interpreted; inspect the control section, detection reagents and section quality (general IHC practice). |
💡Expected ST6GAL1 appearanceCall the section positive when the expected cells show discernible cytoplasmic Golgi-like staining, particularly High signal in hepatocytes or tonsillar germinal center cells (HPA tissue IHC); isolated nuclear, uniform background or staining confined to unexpected cells is suspect (UniProt P15907 location; HPA tissue IHC; general IHC practice).
| Golgi topology | Residues 10–26 form the membrane span; residues 27–406 face the lumen (UniProt P15907 topology). This supports an intracellular Golgi pattern but does not identify the catalog antibody's epitope or specify an antigen-retrieval condition. |
| Secreted form | UniProt reports secretion into body fluid by similarity alongside the membrane-bound Golgi form (UniProt P15907 location). HPA warns that secreted variants can complicate agreement between tissue RNA and protein locations (HPA tissue IHC); interpret staining against cell morphology and controls. |
| Tissue-dependent signal | HPA records High staining in liver hepatocytes and tonsillar germinal center cells, Medium in several other listed populations, and Not detected in selected cells (HPA tissue IHC). Those categories are observed patterns, not universal intensity thresholds for every specimen. |
| Antibody evidence | The supplied catalog antibody, CAB080031, has an Enhanced IHC validation status (HPA antibodies). HPA's tissue profile also carries an Enhanced reliability label with medium RNA–protein consistency (HPA tissue IHC); neither label proves that every deposit in a new section is specific. |
| IF/ICC Q&A: what location should be expected? | A Golgi-associated pattern is biologically plausible from the reported membrane location (UniProt P15907 location), but HPA lists no main ICC-IF location or cell-line images here (HPA subcellular). This record cannot establish an observed IF/ICC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive liver or tonsil section is blank. | The result conflicts with HPA's High staining in hepatocytes or germinal center cells (HPA tissue IHC); a run or detection failure is possible (general IHC practice). | Check that the control contains the expected cells, then verify the primary-antibody step, retrieval used, detection reagents and chromogen development against the applicable IHC-P workflow (general IHC practice). No target-specific retrieval setting is supplied. |
| The section shows widespread faint color with little cellular definition. | Background can mask the cytoplasmic Golgi pattern reported by HPA (HPA tissue IHC); excess detection signal or incomplete washing are general possibilities (general IHC practice). | Compare a no-primary control, assess wash and blocking steps, and review chromogen development before lowering the staining score or changing conditions (general IHC practice). |
| Unexpected cells stain strongly, including cells HPA records as Not detected. | Cross-reactivity or endogenous chromogenic activity may explain the discrepancy (HPA tissue IHC: selected cells Not detected; general IHC practice). HPA's category does not guarantee every individual specimen will be blank. | Confirm cell identity on the counterstained section and inspect no-primary and known-positive controls; if unexpected signal persists only with primary antibody, reassess specificity (general IHC practice). |
| Staining is predominantly nuclear or forms a uniform surface rim. | That distribution conflicts with the reported Golgi membrane location and cytoplasmic Golgi tissue pattern (UniProt P15907 location; HPA tissue IHC). | Review section morphology and control staining, then reassess primary-antibody specificity and detection background before interpreting the compartment as ST6GAL1 (general IHC practice). |
| A Medium-staining tissue looks weaker than the liver or tonsil control. | Different intensity is consistent with the supplied HPA categories: several listed populations are Medium, while hepatocytes and tonsillar germinal center cells are High (HPA tissue IHC). | Score the expected cell population and Golgi-like distribution within that tissue; compare sections processed in the same run rather than imposing one intensity cutoff across tissues (general IHC practice). |
| ICC-IF images do not show a clear Golgi pattern. | HPA supplies no ICC-IF cell-line images or main subcellular location for this record, although UniProt places the membrane-bound form in Golgi stacks (HPA subcellular; UniProt P15907 location). | Treat the IF result as unconfirmed by this payload and use the separate IF/ICC guide for assay-specific controls and interpretation; do not transfer an IHC-P staining result into an IF validation claim. |
Experiment evidence
IHC & IF images showing ST6GAL1 staining
A30620 · IHCImmunohistochemical analysis of paraffin-embedded Human stomach. 1, Antibody was diluted at 1:200(4° overnight). 2, High-pressure and temper
A30620 · IHCImmunohistochemistry analysis of paraffin-embedded human prostate carcinoma tissue, using ST6GAL1 Antibody. The picture on the right is bloc
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000073849-ST6GAL1/tissue/cerebral+cortex#imid_21083667
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000073849-ST6GAL1/tissue/colon#imid_21089569
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000073849-ST6GAL1/tissue/liver#imid_21089578
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000073849-ST6GAL1/tissue/kidney#imid_21089563
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ST6GAL1 IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in hepatocytes). Use adipose tissue as the negative comparison because adipocytes are not detected (HPA: Not detected in adipocytes); any unstained cells within the liver section should lack specific perinuclear Golgi staining, but HPA does not identify a defined internal negative cell population there (UniProt P15907 Golgi localisation; HPA: High in hepatocytes).
Positive control tissue: Liver (Hepatocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ST6GAL1; derive a cell-line control from the positive tissue's cell type (Hepatocytes) and confirm it by RNA or western blot first.
Technical controls: Run no-primary (secondary-only) and concentration-matched isotype controls matched to the primary antibody’s host species and clonality, plus an ST6GAL1 knockout biological control processed in parallel (standard IHC practice). For liver chromogenic IHC, quench endogenous peroxidase and assess endogenous biotin if using avidin-biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A30620 paraffin-section caption does not state a fixative (A30620 tissue-IHC caption). The caption uses high-pressure, high-temperature EDTA retrieval at pH 8.0 and a 1:200 primary dilution overnight at 4°C; it provides a starting condition but does not establish retrieval dependency (A30620 tissue-IHC caption). No supplied comparison establishes whether frozen sections or IF are easier; endogenous peroxidase and, with avidin-biotin detection, endogenous biotin can complicate liver chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for ST6GAL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial 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 → |
| Breast | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ST6GAL1 IHC Tips
Use Golgi-pattern staining and cell-type controls to troubleshoot ST6GAL1 in paraffin-section chromogenic IHC (UniProt P15907; HPA tissue IHC).
Which retrieval condition should I try first for weak ST6GAL1 staining?
Start with heat-mediated retrieval in EDTA pH 8.0 (datasheet A30620). The selected paraffin-section image used high pressure and temperature retrieval, followed by primary antibody at 1:200 overnight at 4°C (datasheet A30620). If staining remains weak, compare a shorter and a longer heating interval on matched sections while keeping detection and incubation conditions constant (standard IHC practice). Assess recovery by looking for a cytoplasmic Golgi pattern in expected positive cells, including hepatocytes, rather than accepting diffuse staining alone (HPA tissue IHC). Record tissue damage and background alongside signal, since excessive heating can compromise section morphology (standard IHC practice).
Could fixation explain weak or uneven ST6GAL1 staining?
Target-specific fixation sensitivity is unknown: the selected image identifies paraffin-embedded human stomach but does not state its fixative (datasheet A30620). Compare sections with documented fixation histories and keep section thickness, retrieval, antibody incubation and chromogen development consistent (standard IHC practice). For the catalog antibody, begin with EDTA pH 8.0 retrieval and 1:200 primary antibody overnight at 4°C (datasheet A30620). If signal varies between blocks, inspect preservation and tissue edges before attributing the difference to ST6GAL1 abundance (standard IHC practice). A convincing positive should retain the expected cytoplasmic Golgi pattern in an appropriate cell population (HPA tissue IHC).
What staining pattern should count as plausible ST6GAL1 localisation?
Look for cytoplasmic staining with a Golgi pattern in tissue cells, as reported for ST6GAL1 (HPA tissue IHC). The membrane-bound protein has a transmembrane segment at residues 10–26, with residues 27–406 on the luminal side (UniProt P15907 topology). Compare suspected staining with hepatocytes or tonsillar germinal center cells, both reported as strongly positive, while keeping exposure to chromogen comparable (HPA tissue IHC; standard IHC practice). A secreted form is also described, so extracellular signal needs careful interpretation rather than automatic rejection (UniProt P15907 subcellular annotation). Predominantly nuclear staining should prompt review of controls and detection background because it does not match the reported Golgi localisation (UniProt P15907; HPA tissue IHC).
How do isoforms and epitope position affect an IHC result?
ST6GAL1 has 2 annotated isoforms, but the supplied record does not map their differences or the catalog antibody’s epitope (UniProt P15907; datasheet A30620). Request epitope mapping before claiming isoform-specific detection, and avoid treating a negative section as evidence that one isoform is absent (standard IHC interpretation). The annotated cytoplasmic segment is residues 1–9, whereas residues 27–406 face the Golgi lumen in the membrane-bound form (UniProt P15907 topology). Glycosylation sites at 149 and 161 warrant caution when interpreting epitope accessibility, although their effect on this antibody is unknown (UniProt P15907). Compare any alternate antibody’s staining pattern against the same positive and negative tissue controls (standard IHC practice).
How should I assess ST6GAL1 in a multiplex IF experiment?
For the separate IF workflow, pair ST6GAL1 with an albumin hepatocyte marker and check whether signal occupies a plausible Golgi region in those cells (HPA tissue IHC; UniProt P15907 subcellular annotation). Choose a fluorophore whose emission is separated from tissue autofluorescence, then inspect unstained and single-label controls before interpreting colocalisation (standard IF practice). The membrane-bound protein spans residues 10–26; its short cytoplasmic tail and long luminal portion make epitope position relevant to permeabilisation (UniProt P15907 topology). Because this antibody’s epitope and IF performance are unspecified, establish permeabilisation empirically using mapped epitope information and matched controls (datasheet A30620; standard IF practice). The paraffin-section 1:200 dilution is IHC evidence, not an IF dilution recommendation (datasheet A30620).
How can I reduce diffuse brown staining without losing Golgi signal?
Run a no-primary section and an appropriate negative tissue control to separate detection background from antibody-dependent staining (standard IHC practice). Adipocytes and cardiomyocytes are reported as not detected, while hepatocytes show high staining, providing useful comparison populations when present in the study (HPA tissue IHC). Include a peroxidase block before chromogenic detection and control DAB development time across sections (standard IHC practice). If background persists, titrate around the documented 1:200 primary dilution while retaining the specified EDTA pH 8.0 retrieval as the starting condition (datasheet A30620; standard IHC practice). Prioritise a crisp cytoplasmic Golgi pattern over uniformly stronger brown signal (HPA tissue IHC).
How should I score ST6GAL1 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and staining compartment before scoring; ST6GAL1 has a reported cytoplasmic Golgi pattern in tissues (HPA tissue IHC). For comparable regions, report the percentage of positive cells and an H-score based on intensity categories, or count positive cells per mm² when density is the question (standard IHC practice). Normalise cell counts to viable tissue area and compare like cell types, since hepatocytes and tonsillar germinal center cells are reported as highly positive (HPA tissue IHC). Keep retrieval, the documented 1:200 primary dilution and chromogen development consistent across a scoring batch (datasheet A30620; standard IHC practice). Record extracellular staining separately because a secreted form is described (UniProt P15907 subcellular annotation).
What distinguishes a true ST6GAL1 positive from an artefact?
A credible positive shows a cytoplasmic Golgi pattern in an expected cell population, such as hepatocytes or tonsillar germinal center cells (HPA tissue IHC). Check suspicious nuclear or widespread uniform staining against no-primary controls and tissue regions reported as not detected, such as adipocytes (UniProt P15907; HPA tissue IHC; standard IHC practice). Exclude staining confined to section edges, necrotic areas or sites of residual endogenous peroxidase before assigning a positive score (standard IHC practice). Extracellular staining deserves separate review because ST6GAL1 also has a reported secreted form (UniProt P15907 subcellular annotation). Interpret staining and RNA cautiously: tissue-level agreement has medium consistency, and secretion can complicate their spatial correlation (HPA tissue IHC).
Boster reagents
Best ST6GAL1 / Beta-galactoside alpha-2,6-sialyltransferase 1 IHC Antibodies
The anti-ST6GAL1 catalog antibody has real IHC images from human stomach and prostate carcinoma paraffin sections (catalog IHC captions). IF/ICC is listed, but no IF image is supplied (catalog applications; IF image payload).
Real IHC data
Anti-ST6GAL1/Cd75 Antibody
Cat # A30620
A30620 has IHC images from human stomach and prostate carcinoma paraffin sections (catalog IHC captions). Its listed applications include IHC, IF and ICC, and its listed reactivity covers human, mouse and rat (catalog applications; catalog reactivity).
Which to pick: Choose A30620 for paraffin-section IHC: its human stomach image reports 1:200 antibody dilution and EDTA retrieval at pH 8.0; the fixative is unreported (catalog stomach IHC caption). A30620 is also the listed IF/ICC option, although no IF image is supplied (catalog applications; IF image payload). For mouse or rat work, A30620 lists reactivity with both species, but the supplied tissue images are human; it is polyclonal (catalog reactivity; catalog IHC captions; catalog dilution record).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P15907 (SIAT1_HUMAN, Beta-galactoside alpha-2,6-sialyltransferase 1).
- Human Protein Atlas. ST6GAL1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ST6GAL1 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. ST6GAL1 antibody validation summary (1 antibodies).
- Epigenetic inactivation of ST6GAL1 in human bladder cancer. BMC cancer 2014 — PMC4265431.
- Transcriptomic and metabolomic insights into the variety of sperm storage in oviduct of egg layers. Poultry science 2021 — PMC8082553.
- Extracellular sialyltransferase st6gal1 in breast tumor cell growth and invasiveness. Cancer gene therapy 2022 — PMC9663294.
- Galectin-1-mediated cell adhesion, invasion and cell death in human anaplastic large cell lymphoma: regulatory roles of cell surface glycans. International journal of oncology 2014 — PMC4027875.
- PubMed PMID:2408023 — UniProt-cited evidence.
- PubMed PMID:2373995 — UniProt-cited evidence.
- PubMed PMID:1730763 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ST6GAL1 (Beta-galactoside alpha-2,6-sialyltransferase 1). Expected localisation Cytoplasmic Golgi pattern (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A30620). Validated IHC antibody: A30620 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human stomach tissue; fixative not specified (datasheet A30620); 2. Fixation: Image fixative and duration unreported (datasheet A30620); 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 A30620); 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-ST6GAL1, 1:100 - 1:300 (datasheet A30620); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30620); Caveat: A secreted variant may alter tissue staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; cytoplasmic versus luminal epitope coverage is unknown (UniProt); Regulation: Liver and lymphoid tissue group enrichment (HPA tissue IHC). Evidence: UniProt P15907, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A30620, PMC9663294, PMC4027875. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ST6GAL1/immunohistochemistry-ihc-guide.