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ST3GAL3 / CMP-N-acetylneuraminate-beta-1,4-galactoside alpha-2,3-sialyltransferase · IHC design guide
Design Immunohistochemistry for ST3GAL3
Plan chromogenic IHC for ST3GAL3 using the cytoplasmic tissue staining profile (HPA tissue IHC) and Golgi localization as a molecular expectation (UniProt). Compare high-staining liver hepatocytes with adipocytes reported as undetected, while treating the tissue staining profile as pending external verification (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ST3GAL3 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across most tissue types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+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 A08054-1) | |
| Caveat | A secreted form may complicate cellular staining (UniProt) | |
| Regulation | Expression varies by tissue and cell type (HPA tissue IHC) | |
| Isoform / epitope | 26 isoforms; check cytoplasmic vs lumenal epitope coverage (UniProt) |
Section 1
Recommended ST3GAL3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published ST3GAL3 gastric biopsy protocol (PMC9577379).
| Sample | Paraffin-embedded human liver damage tissue; fixative not specified (datasheet A08054-1) |
| Fixation | Image fixative and duration unreported (datasheet A08054-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in 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-ST3GAL3, 1:50-1:200 (datasheet A08054-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ST3GAL3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); the published protocol used microwave citrate retrieval for 15 min (PMC9577379).
Section 2
What Is the Expected ST3GAL3 Staining Pattern?
ST3GAL3 is a Golgi stack membrane protein with a short cytoplasmic tail and a lumenal catalytic region; a secreted form is also reported (UniProt Q11203). In tissue IHC, expect predominantly cytoplasmic staining in selected glandular cells, hepatocytes, germinal center cells, and pancreatic endocrine cells (HPA tissue IHC). Treat these expectations as provisional: HPA rates its tissue IHC evidence Uncertain, pending external verification (HPA tissue IHC).
| Cytoplasmic staining in appendix or duodenal glandular cells, liver hepatocytes, lymph node germinal center cells, or pancreatic endocrine cells (HPA tissue IHC). | These cell populations are scored High by HPA and provide provisional positive comparators (HPA tissue IHC). A cytoplasmic pattern is compatible with Golgi localization, although routine chromogenic IHC cannot establish the stained organelle on its own (UniProt Q11203; standard IHC interpretation). |
| Predominantly nuclear staining, or an isolated crisp plasma membrane outline without convincing cytoplasmic staining. | Neither pattern is the expected tissue IHC profile (HPA tissue IHC). UniProt places the membrane bound form in the Golgi stack, so check whether the signal is reproducible and antibody dependent before assigning it to ST3GAL3 (UniProt Q11203 topology; standard IHC interpretation). |
| Strong signal in adipocytes, skeletal myocytes, or placental trophoblastic cells, especially when the expected positive cells are faint. | HPA reports these cells as Not detected in its tissue IHC dataset (HPA tissue IHC). Investigate antibody cross reactivity or endogenous detection activity; skeletal muscle needs particular caution because UniProt reports high adult expression there (UniProt Q11203 tissue specificity). |
| Diffuse color across multiple cell types, extracellular spaces, and the section background. | A widespread haze does not resemble HPA's cell selective cytoplasmic profile (HPA tissue IHC). Assess nonspecific antibody binding, incomplete blocking, and endogenous detection activity with appropriate control sections (standard chromogenic IHC practice). |
| No convincing signal in a selected HPA high cell population while tissue morphology and counterstain remain interpretable. | Check the staining run before calling the sample ST3GAL3 negative: the HPA high designation is provisional, and its antibody HPA051102 has Uncertain IHC validation (HPA tissue IHC; HPA antibodies). A second positive tissue can help distinguish a sample result from a failed run (standard IHC practice). |
💡Expected ST3GAL3 appearanceCall a result provisionally positive when selected HPA High cell populations show cytoplasmic staining above adjacent background; isolated nuclear signal or similarly strong staining throughout HPA Not detected populations warrants investigation (HPA tissue IHC; UniProt Q11203 localization).
| Golgi topology and interpretation | Residues 9–28 form a transmembrane segment, with residues 29–375 lumenal (UniProt Q11203 topology). This supports an intracellular membrane associated expectation, but a chromogenic cytoplasmic signal alone does not resolve Golgi cisternae (standard IHC interpretation). |
| Tissue and cell choice | HPA scores glandular cells in adrenal gland, appendix, duodenum, parathyroid gland, and seminal vesicle High; it also scores hepatocytes, germinal center cells, and pancreatic endocrine cells High (HPA tissue IHC). Select the scored cell population, not the entire organ, for comparison. |
| Evidence disagreement | UniProt reports high expression in adult skeletal muscle, while HPA scores skeletal myocytes Not detected by tissue IHC (UniProt Q11203 tissue specificity; HPA tissue IHC). Interpret a negative muscle section as assay specific evidence, not proof that the protein is absent. |
| Processing and molecular diversity | UniProt reports a secreted form, two glycosylation sites at residues 80 and 171, and 26 isoforms (UniProt Q11203). Without an antibody epitope or isoform coverage record, these annotations cannot predict which forms a particular IHC stain detects. |
| Validation strength | HPA describes its tissue IHC reliability as Uncertain, pending external verification, and lists HPA051102 IHC as Uncertain (HPA tissue IHC; HPA antibodies). Use matched positive and negative comparators when judging a new staining pattern (standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| A selected HPA high population is blank (HPA tissue IHC). | A failed staining run, low effective antibody signal, or an unsupported epitope assumption could explain the result (standard IHC practice). HPA's high score is provisional (HPA tissue IHC). | Check the run with another HPA high population, then review the catalog antibody's IHC-P instructions and the run's antigen retrieval, dilution, and detection controls (standard IHC practice). |
| Nuclei dominate the stain. | Nuclear localization does not match the reported cytoplasmic tissue profile or Golgi membrane localization (HPA tissue IHC; UniProt Q11203). | Compare with a no primary control and a separate positive tissue; reassess antibody specificity if nuclear staining persists (standard IHC practice). |
| HPA not detected cell types stain as strongly as positive comparators (HPA tissue IHC). | Cross reactivity or endogenous chromogenic detection activity is plausible (standard IHC practice); HPA's tissue calls remain uncertain (HPA tissue IHC). | Check no primary and detection controls, compare cell types within the same section, and seek independent antibody evidence before reclassifying the pattern (standard IHC practice). |
| The whole section has diffuse color. | Nonspecific binding or residual endogenous detection activity can obscure a cell selective cytoplasmic pattern (standard IHC practice; HPA tissue IHC). | Review blocking, washes, antibody dilution, and detection controls; retain a counterstain that lets the scored cell population be identified (standard chromogenic IHC practice). |
| Skeletal muscle is negative while a liver comparator is positive. | This resembles HPA's myocyte Not detected and hepatocyte High IHC scores, despite UniProt's adult skeletal muscle expression report (HPA tissue IHC; UniProt Q11203). | Report the disagreement with the assay and cell type specified; avoid treating either source as a validated sensitivity test for the other (HPA tissue IHC; UniProt Q11203). |
| Can IF/ICC confirm the subcellular pattern? | HPA summarizes ST3GAL3 as membrane localized but provides no ICC-IF images or main location call; UniProt places the membrane bound form in Golgi trans cisternae (HPA subcellular; UniProt Q11203). | Treat IF/ICC localization as a separate validation question. Compare any result with a Golgi reference and appropriate controls; do not infer an IF protocol or confirmed image pattern from these records (standard IF practice; HPA subcellular). |
Experiment evidence
IHC & IF images showing ST3GAL3 staining
A08054-1 · IHCImmunohistochemistry of paraffin-embedded human liver damage using ST3GAL3 antibody at dilution of 1:100 .Perform microwave antigen retrieva
A08054-1 · IHCImmunohistochemistry of paraffin-embedded rat lung using ST3GAL3 antibody at dilution of 1:100 .Perform microwave antigen retrieval with 10
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000126091-ST3GAL3/tissue/cerebral+cortex#imid_14099782
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000126091-ST3GAL3/tissue/colon#imid_14099708
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000126091-ST3GAL3/tissue/liver#imid_14099709
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000126091-ST3GAL3/tissue/kidney#imid_14099710
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ST3GAL3 IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in liver hepatocytes). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected in oral mucosa squamous epithelial cells); on the liver slide, treat nonhepatocyte cells as internal negatives only where they are visibly unstained.
Positive control tissue: Adrenal gland (Glandular cells, 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 ST3GAL3; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and ST3GAL3 knockout tissue as a biological negative. For chromogenic liver IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A08054-1 liver caption does not state its fixative (selected-SKU tissue-IHC caption). That caption uses paraffin sections, microwave retrieval in 10 mM PBS at pH 7.2, and a 1:100 antibody dilution (selected-SKU tissue-IHC caption); whether retrieval is required across specimens is unreported. Frozen sections and IF cannot be judged easier from the supplied evidence, and no ICC-IF image-bearing cell lines are listed (HPA subcellular record).
HPA tissue IHC evidence for ST3GAL3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Pending external verification.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ST3GAL3 IHC Tips
Troubleshooting paraffin section chromogenic IHC for ST3GAL3, with one question on adapting the assay to IF.
What retrieval conditions should I try first for ST3GAL3 in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval protocol). Let sections cool in buffer, then compare staining with a matched positive control while keeping antibody dilution and detection conditions constant (standard IHC practice). If staining is weak, test microwave retrieval in 10 mM PBS at pH 7.2 as a fallback; that condition was used with the catalog antibody on paraffin embedded human liver damage at 1:100 (A08054-1 image caption). Record tissue morphology alongside signal, since excessive heating can damage sections and complicate interpretation (standard IHC practice).
How should I troubleshoot weak staining when section fixation is uncertain?
The selected paraffin section caption does not state a fixative, so target specific fixation sensitivity is unknown (A08054-1 image caption). Record the fixative and processing history for each specimen, then compare sections processed the same way before changing antibody or detection conditions (standard IHC practice). Check that deparaffinisation is complete and that the pH 6.0, 95–98 °C, 20 min retrieval step was performed consistently (page retrieval protocol; standard IHC practice). If staining remains weak, compare a separately processed control section and report its fixation history; neither the tissue expression profile nor Golgi topology establishes how fixation affects this antibody (HPA tissue IHC; UniProt Q11203 topology).
Where should convincing ST3GAL3 staining appear within positive cells?
Assess chromogenic staining primarily within the cell body, with a compact perinuclear pattern being plausible for a Golgi stack membrane protein (UniProt Q11203 subcellular location; standard IHC interpretation). ST3GAL3 has a membrane spanning segment at residues 9–28, a short cytoplasmic segment at 1–8, and a lumenal region at 29–375 (UniProt Q11203 topology). HPA describes cytoplasmic staining in most tissues, but its tissue IHC reliability is uncertain pending external verification (HPA tissue IHC). Compare the suspected signal with adjacent cells and section controls; uniform nuclear or tissue edge staining warrants an artefact check before assigning it to ST3GAL3 (standard IHC practice).
Could isoforms or epitope position explain variable IHC staining?
ST3GAL3 has 26 listed isoforms, so an antibody’s recognised sequence matters when comparing sections or studies (UniProt Q11203 isoforms). Locate the catalog antibody’s stated immunogen or epitope before claiming that it detects every isoform; no epitope position is supplied here (catalog antibody evidence supplied; standard antibody validation practice). The reference protein has a cytoplasmic segment at residues 1–8, a membrane segment at 9–28, and a lumenal region at 29–375, with glycosylation sites at 80 and 171 (UniProt Q11203 topology and glycosylation). Compare staining with an independently validated reagent or orthogonal assay when isoform coverage or epitope accessibility changes the biological conclusion (standard IHC validation practice).
How can I adapt this IHC result to multiplex ST3GAL3 immunofluorescence?
Treat the chromogenic paraffin section result as IHC evidence, then establish IF staining and controls separately for this antibody (A08054-1 image caption; standard IF validation practice). Multiplex ST3GAL3 with a validated marker for the expected cell type, such as hepatocytes in a liver section, and confirm that the signals occupy the same cells (HPA: High in hepatocytes; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and include single stain and no primary controls when setting exposure and bleed through limits (standard IF practice). For a cytoplasmic epitope, test mild permeabilisation; for a lumenal epitope within residues 29–375, verify that permeabilisation permits access without disrupting the Golgi pattern (UniProt Q11203 topology; standard IF practice).
What should I change when the DAB signal is diffuse or widespread?
Check a no primary control and examine whether brown signal follows tissue edges, damaged regions, or every cell indiscriminately (standard chromogenic IHC practice). Block endogenous peroxidase before DAB development, then adjust primary antibody concentration and detection incubation one variable at a time using matched sections (standard chromogenic IHC practice). The catalog image used antibody at 1:100 in paraffin embedded human liver damage, which is a documented starting point rather than a validated dilution for every specimen (A08054-1 image caption). Compare the remaining cellular signal with the expected Golgi associated location and with control tissue, while recognising the uncertain reliability of HPA tissue staining (UniProt Q11203 subcellular location; HPA tissue IHC).
How should I quantify ST3GAL3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis region before scoring, and apply the same DAB threshold and imaging settings to every section (standard quantitative IHC practice). Report the percentage of positive cells and, when intensity is reproducible, an H-score using percentages at intensity grades 0–3, giving a range of 0–300 (standard IHC scoring practice). For sparse positive cells, report positive cell density per mm² of evaluable tissue and exclude folds, edges, and necrosis using fixed criteria (standard quantitative IHC practice). Normalise counts to the number of eligible cells or evaluable tissue area, and retain compartment and cell type annotations because ST3GAL3 is Golgi associated and HPA patterns vary by cell type (UniProt Q11203 subcellular location; HPA tissue IHC).
When is apparent ST3GAL3 positivity more likely to be artefact?
Give most weight to cellular staining compatible with a Golgi associated enzyme and to a reproducible pattern in the intended cell population (UniProt Q11203 subcellular location; standard IHC interpretation). Question isolated nuclear staining, abrupt signal restricted to section edges, necrotic debris, or color that persists without primary antibody (standard chromogenic IHC practice). In liver, hepatocyte positivity is reported by HPA, while its tissue IHC reliability remains uncertain pending external verification (HPA: High in hepatocytes; HPA tissue IHC reliability). Resolve disagreements with matched positive and no primary controls, review endogenous peroxidase blocking, and avoid calling secretion or enzyme activity from DAB localisation alone (standard IHC practice; UniProt Q11203 subcellular location and function).
Boster reagents
Best ST3GAL3 / CMP-N-acetylneuraminate-beta-1,4-galactoside alpha-2,3-sialyltransferase IHC Antibodies
A08054-1 has paraffin-section IHC images from human liver damage, rat lung, and mouse spinal cord (catalog IHC captions). IF/ICC is listed, but no IF image is supplied (catalog applications and image alts).
Real IHC data
Anti-ST3GAL3 Antibody
Cat # A08054-1
A08054-1 has IHC images of paraffin-embedded human liver damage, rat lung, and mouse spinal cord (catalog IHC captions). A08054-1 lists IF/ICC and Human, Mouse, and Rat reactivity; no IF image is supplied (catalog applications, reactivity, and image alts).
Which to pick: Choose A08054-1 for paraffin-section IHC: its captions document 1:100 staining after microwave retrieval in 10 mM PBS, pH 7.2; the fixative is unreported (catalog IHC captions). For IF/ICC, A08054-1 lists those applications and a 1:50–1:200 dilution, but provides no IF image; clonality is unreported (catalog applications, IF dilution, image alts, and clone field). For work across human, mouse, and rat, A08054-1 lists reactivity with all three and has a paraffin-section IHC caption for each (catalog reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q11203 (SIAT6_HUMAN, CMP-N-acetylneuraminate-beta-1,4-galactoside alpha-2,3-sialyltransferase).
- Human Protein Atlas. ST3GAL3 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ST3GAL3 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. ST3GAL3 antibody validation summary (1 antibodies).
- alpha2,3 sialic acid processing enzymes expression in gastric cancer tissues reveals that ST3Gal3 but not Neu3 are associated with Lauren's classification, angiolymphatic invasion and histological grade. European journal of histochemistry : EJH 2022 — PMC9577379.
- Sialyltransferase Mutations Alter the Expression of Calcium-Binding Interneurons in Mice Neocortex, Hippocampus and Striatum. International journal of molecular sciences 2023 — PMC10743413.
- PubMed PMID:8333853 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ST3GAL3 (CMP-N-acetylneuraminate-beta-1,4-galactoside alpha-2,3-sialyltransferase). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08054-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver damage tissue; fixative not specified (datasheet A08054-1); 2. Fixation: Image fixative and duration unreported (datasheet A08054-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in 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-ST3GAL3, 1:50-1:200 (datasheet A08054-1); 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 A08054-1); Caveat: A secreted form may complicate cellular staining (UniProt); Isoform / epitope: 26 isoforms; check cytoplasmic vs lumenal epitope coverage (UniProt); Regulation: Expression varies by tissue and cell type (HPA tissue IHC). Evidence: UniProt Q11203, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A08054-1, PMC9577379. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ST3GAL3/immunohistochemistry-ihc-guide.