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B4GALT2 / Beta-1,4-galactosyltransferase 2 · IHC design guide
Design Immunohistochemistry for B4GALT2
Plan B4GALT2 paraffin-section IHC around granular tissue staining (HPA tissue IHC) and an expected Golgi membrane location (UniProt). Start with the catalog antibody’s 1:100–1:300 IHC dilution range (datasheet A10774T2), using glandular cells in colon as a positive reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
B4GALT2 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 | Golgi stack membrane expected; tissue compartment unverified (UniProt) | |
| Staining pattern | Granular staining in several tissues, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10774T2) | |
| 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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining may confound interpretation (HPA tissue IHC) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope maps unknown; cytosolic versus lumenal location matters (UniProt) |
Section 1
Recommended B4GALT2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A10774T2) is accompanied by one published lung adenocarcinoma IHC protocol (PMC11865799).
| Sample | Paraffin-embedded human cervical carcinoma tissue; fixative not specified (datasheet A10774T2) |
| Fixation | Image fixative and duration unreported (datasheet A10774T2); 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 A10774T2); 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-B4GALT2, 1:100-1:300 (datasheet A10774T2) |
| 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 | B4GALT2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 (datasheet A10774T2); the published protocol does not specify retrieval conditions (PMC11865799).
Section 2
What Is the Expected B4GALT2 Staining Pattern?
B4GALT2 is a Golgi stack membrane protein with a cytoplasmic N terminus and lumenal catalytic region (UniProt O60909 topology). Expect granular cytoplasmic staining in the appropriate cells, including colon and duodenal glandular cells (HPA: High in both). HPA rates tissue IHC reliability Supported, with medium agreement between staining and RNA data; presumed off-target staining was observed and disregarded (HPA: tissue IHC reliability).
| Granular cytoplasmic staining in colon or duodenal glandular cells (HPA: High in both cell types). | Consistent with the reported tissue pattern and Golgi localization (HPA: tissue IHC; UniProt O60909 subcellular location). Score the relevant cells and their staining intensity; a positive region does not imply that every cell in the section is positive. |
| Predominantly nuclear or continuous plasma membrane staining, with little granular cytoplasmic signal. | This differs from the Golgi pattern expected from UniProt and HPA (UniProt O60909 subcellular location; HPA: subcellular location). Treat it as suspect and compare a known-positive tissue section and detection controls before assigning B4GALT2 expression. |
| Strong staining in adipocytes or esophageal squamous epithelial cells (HPA: Not detected in these cells). | These are unexpected cell-specific results, even if another cell population in the tissue stains. Consider antibody cross-reactivity or chromogenic detection activity; HPA reports presumed off-target staining in its tissue assessment (HPA: tissue IHC reliability). |
| Uniform haze across cells, extracellular spaces, or the whole section. | A broad background pattern obscures the cell-restricted, granular pattern reported by HPA (HPA: tissue IHC profile). Compare a no-primary control, review blocking and washes, and avoid scoring haze as B4GALT2 staining. |
| No signal in colon glandular cells on a section expected to be positive (HPA: High in colon glandular cells). | The result is inconclusive until the staining run is checked. Review tissue integrity, antigen retrieval, primary antibody use and chromogenic detection; the HPA tissue record does not identify a B4GALT2-specific cause of signal loss. |
💡Expected B4GALT2 appearanceCall a positive IHC result when the appropriate glandular or other HPA-positive cells show clear granular cytoplasmic staining (HPA: tissue IHC profile; UniProt O60909 Golgi location); diffuse haze, dominant nuclear staining, or strong staining in HPA-undetected cell types warrants review.
| Golgi membrane topology (UniProt O60909 topology) | B4GALT2 has one membrane segment at residues 16–36; residues 1–15 face the cytoplasm and 37–372 face the lumen (UniProt O60909 topology). This supports an intracellular Golgi pattern, but does not predict how an unspecified antibody epitope responds to antigen retrieval. |
| Cell type within a tissue (HPA: tissue IHC) | Interpret staining at the cell level. HPA reports High staining in fallopian tube ciliated cell bodies and Not detected in ovarian stroma cells (HPA: tissue IHC). The latter does not make the entire ovary a negative control; UniProt reports relatively high expression in ovary (UniProt O60909 tissue specificity). |
| Antibody evidence (HPA: antibody validation) | The listed IHC antibody HPA047739 has Supported IHC validation; HPA007620 has Supported ICC validation, with no IHC status listed (HPA: antibody validation). HPA also notes presumed off-target tissue staining, so evaluate unexpected patterns against cell-specific references (HPA: tissue IHC reliability). |
| IF/ICC: what localization should be expected? | HPA reports mainly Golgi apparatus and vesicle localization in ICC-IF, with Golgi supported and vesicles approved (HPA: subcellular). This informs image interpretation; the ICC-IF record does not establish an IHC-P staining protocol or a dilution. |
| Isoforms and glycosylation (UniProt O60909) | UniProt lists three isoforms and glycosylation sites at residues 66, 71 and 357 (UniProt O60909). Without an antibody epitope or isoform-specific staining data, these annotations cannot establish which isoforms an IHC signal detects or explain an unexpected tissue result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive glandular cells have no visible signal (HPA: High in colon glandular cells). | A staining-run failure is possible; the supplied sources do not establish B4GALT2-specific fixation sensitivity. | Check a positive control section, reagent application, retrieval settings and detection steps. Repeat only after identifying a plausible run-level issue; do not attribute the loss to fixation from these records. |
| Staining is broad and diffuse instead of granular (HPA: granular tissue expression). | Nonspecific antibody binding or detection background may obscure the expected pattern. | Inspect the no-primary control, blocking, washes and primary concentration. Compare cell boundaries and unstained regions before assigning a positive score. |
| Adipocytes stain strongly (HPA: Not detected in adipocytes). | Cross-reactivity or endogenous detection activity is possible; HPA reports presumed off-target tissue staining (HPA: tissue IHC reliability). | Compare no-primary and known-positive sections, then reassess whether the signal is cell-specific and granular. Do not treat adipocyte staining alone as confirmation. |
| Staining appears mainly nuclear or along the cell surface. | The compartment conflicts with the Golgi location reported by UniProt and HPA (UniProt O60909; HPA: subcellular). | Review morphology, counterstain and detection controls. Confirm a granular intracellular pattern in a known-positive cell type before interpreting the unusual compartment. |
| Ovarian stroma is unstained, but other ovarian cells stain. | HPA reports Not detected specifically in ovarian stroma cells; that observation does not describe every ovarian cell (HPA: tissue IHC). | Score the cell populations separately. Compare the stained population with tissue morphology and avoid calling the entire section either positive or negative from stromal staining alone. |
| IF/ICC shows vesicle signal in addition to Golgi signal. | HPA lists vesicles as an approved location and Golgi as supported (HPA: subcellular). | Describe each compartment separately and use the IF/ICC guide for that application. Do not transfer ICC-IF validation or settings to paraffin-section IHC (HPA: antibody validation). |
Experiment evidence
IHC & IF images showing B4GALT2 staining
A10774T2 · IHCImmunohistochemical analysis of paraffin-embedded human cervical carcinoma. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, p
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117411-B4GALT2/tissue/cerebral+cortex#imid_14094223
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117411-B4GALT2/tissue/colon#imid_14096372
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117411-B4GALT2/tissue/liver#imid_14096373
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000117411-B4GALT2/tissue/kidney#imid_14096374
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA007620; green = B4GALT2, blue = nucleus, red = microtubules.
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA007620; green = B4GALT2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for B4GALT2 IHC & IF
🧪Run colon first: glandular cells should stain strongly (HPA: colon glandular cells High). Use esophageal squamous epithelium as the negative tissue (HPA: esophagus squamous epithelial cells Not detected); compare non-glandular cells on the colon slide for background, while treating their B4GALT2 status as unverified by the supplied HPA row.
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 ICC-IF images show B4GALT2 in PC-3, U-251MG, U2OS, with annotated localisation: Golgi apparatus (supported), Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus B4GALT2 knockout tissue as a biological negative. Quench endogenous peroxidase and inspect inflammatory cells for residual signal in colon (standard chromogenic IHC practice).
⚠️Feasibility: No supplied source reports a B4GALT2-specific fixation window or fixation effect, and the selected paraffin-section caption does not state its fixative (A10774T2 caption: fixative unreported). Start IHC-P optimization with Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 (A10774T2 caption). The evidence does not establish that frozen sections or IF are easier; ICC-IF images document Golgi and vesicle localization (HPA subcellular: PC-3, U-251MG, U2OS), while colonic inflammatory cells can contribute endogenous peroxidase signal (standard IHC practice).
HPA tissue IHC evidence for B4GALT2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced B4GALT2 IHC Tips
Troubleshoot B4GALT2 staining in paraffin sections by checking retrieval, cellular pattern, controls, and cell type before scoring chromogenic signal.
Which retrieval conditions should I try when B4GALT2 staining is weak?
Use heat-mediated Tris-EDTA at pH 9.0 for B4GALT2 staining in paraffin sections (datasheet A10774T2). The selected antibody caption pairs this retrieval with a 1:200 primary dilution incubated at 4 °C overnight, but does not report the fixative or heating duration (caption A10774T2). If signal remains weak, compare retrieval heating times on matched sections while keeping antibody dilution and detection constant, and inspect morphology for heat damage (standard IHC practice). Only after checking those conditions, consider citrate at pH 6.0 as a fallback experiment, using a positive control and treating the result as a new optimisation rather than a documented protocol (standard IHC practice).
How should I troubleshoot variable staining across differently fixed specimens?
B4GALT2-specific fixation sensitivity is unknown: the selected image caption says paraffin-embedded but does not identify a fixative (caption A10774T2). Record each specimen’s fixative and fixation interval, then compare similarly processed sections with the same Tris-EDTA retrieval at pH 9.0 and the same 1:200 primary dilution (datasheet A10774T2; standard IHC practice). A controlled pilot comparing, for example, 6 and 24 hours of fixation can test whether processing contributes to signal loss; those times are proposed test conditions, not B4GALT2 findings (standard IHC practice). Include a morphology check and a matched positive-control section, because changes in staining alone cannot identify which processing step caused the difference (standard IHC practice).
What staining pattern is plausible for B4GALT2 in tissue sections?
Expect predominantly granular, perinuclear cytoplasmic staining consistent with a Golgi-associated protein, rather than uniform nuclear staining (UniProt O60909 localisation; HPA tissue IHC profile). B4GALT2 is assigned to the trans cisternae of the Golgi stack, with a membrane span at residues 16–36 and a lumenal region at 37–372 (UniProt O60909 topology). HPA reports granular staining in several tissues and supported Golgi localisation with an approved vesicular location in cell imaging (HPA tissue IHC; HPA subcellular). Assess the pattern within identified cells at high magnification, and compare granular signal with a no-primary control before calling diffuse cytoplasmic colour specific (standard IHC practice).
Could epitope location or isoform choice explain discordant staining?
B4GALT2 has 3 annotated isoforms, so recognition across variants cannot be assumed without the antibody’s epitope information (UniProt O60909 isoforms; standard antibody validation practice). Map the stated immunogen, if available, against the 1–15 cytoplasmic segment, 16–36 membrane span, and 37–372 lumenal region before interpreting a negative section (UniProt O60909 topology). The protein has annotated glycosylation sites at residues 66, 71, and 357, but the supplied evidence does not establish whether they affect this antibody’s tissue staining (UniProt O60909 glycosylation; caption A10774T2). Compare matched positive-control sections under the documented pH 9.0 retrieval condition and seek antibody-specific epitope data before assigning an isoform-specific result (datasheet A10774T2; standard IHC practice).
How can IF help assess the IHC pattern without misreading Golgi signal?
For a separate IF experiment, pair B4GALT2 with a glandular epithelial marker when examining a tissue compartment reported to stain strongly, such as colon glandular cells (HPA tissue IHC; standard IF practice). Choose a spectrally separated far-red fluorophore for B4GALT2 and inspect single-label controls where tissue autofluorescence is prominent (standard IF practice). Plan permeabilisation around the antibody’s known epitope: the 1–15 cytoplasmic tail faces cytosol, whereas residues 37–372 face the Golgi lumen and may require access across organelle membranes (UniProt O60909 topology; standard IF practice). Look for perinuclear Golgi signal and possible vesicles, using compartment markers and controls to assess overlap rather than treating colour coincidence as proof of identity (HPA subcellular; standard IF practice).
What should I check when chromogenic background obscures granular staining?
Start with a no-primary section and inspect whether colour persists in the same cells or tissue structures; this helps separate antibody-dependent staining from detection background (standard IHC practice). Use a protein block for nonspecific binding and a peroxidase block before DAB development when using a peroxidase-based detection system (standard IHC practice). The selected caption uses a 1:200 primary dilution at 4 °C overnight and a 1:200 secondary dilution for 45 minutes at room temperature; change one variable at a time when optimising (caption A10774T2; standard IHC practice). HPA explicitly notes presumed off-target binding in its tissue assessment, so require an appropriate cellular pattern and controls before interpreting persistent granular colour as B4GALT2 (HPA tissue IHC reliability description).
How should I score B4GALT2 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring region before reviewing outcome groups, and score granular cytoplasmic signal rather than unrelated nuclear colour (UniProt O60909 localisation; standard IHC practice). An H-score combines the percentage of cells at intensities 0–3 into a 0–300 scale; report the percentage positive alongside it so shifts in prevalence remain visible (standard IHC practice). For focal infiltrates, positive-cell density per mm² can be useful, provided the denominator is viable, evaluable tissue area and the counted cell type is stated (standard IHC practice). Normalise comparisons to the same cell compartment, section quality, staining batch, and control performance because HPA reports low tissue specificity at the RNA level and variable tissue-cell staining (HPA tissue IHC; standard IHC practice).
Which findings support true B4GALT2 staining rather than artefact?
A credible positive shows granular cytoplasmic staining in an appropriate cell population, consistent with Golgi localisation and the reported tissue profile (UniProt O60909 localisation; HPA tissue IHC). Colon glandular cells are reported high, whereas adipocytes are reported not detected; use these as contextual comparisons, accounting for the specific cells present on each slide (HPA tissue IHC). Diffuse nuclear colour, signal confined to section edges or necrotic regions, and staining that remains in a no-primary control warrant investigation before calling positivity (standard IHC practice). Check the peroxidase block when endogenous enzyme activity is suspected, and interpret any isolated positive result cautiously because HPA reports medium staining–RNA consistency and disregarded presumed off-target binding (standard IHC practice; HPA tissue IHC reliability description).
Boster reagents
Best B4GALT2 / Beta-1,4-galactosyltransferase 2 IHC Antibodies
A10774T2 has an IHC image of human paraffin-embedded cervical carcinoma (catalog IHC image caption); IF and human/mouse reactivity are listed, but no IF image is supplied (catalog applications/reactivity; IF image alts).
Real IHC data
Anti-beta-1,4-Gal-T2 B4GALT2 Antibody
Cat # A10774T2
A10774T2 is shown staining paraffin-embedded human cervical carcinoma by IHC (catalog IHC image caption). It lists IHC and IF applications and human/mouse reactivity; no IF image is supplied (catalog applications/reactivity; IF image alts).
Which to pick: For tissue IHC, choose A10774T2: its image caption documents paraffin-embedded human cervical carcinoma, 1:200 primary antibody dilution and Tris-EDTA retrieval at pH 9.0; the fixative is unreported (catalog IHC image caption). For IF, A10774T2 lists 1:50, but supplies no IF image or ICC application entry (catalog IF dilution/applications; IF image alts). For cross-species work, A10774T2 lists human and mouse reactivity, although its IHC image shows human tissue only; rabbit host is listed and clonality is unreported (catalog reactivity/host/clone; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O60909 (B4GT2_HUMAN, Beta-1,4-galactosyltransferase 2).
- Human Protein Atlas. B4GALT2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. B4GALT2 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus and vesicles..
- Human Protein Atlas. B4GALT2 antibody validation summary (2 antibodies).
- Novel post-translational modification learning signature reveals B4GALT2 as an immune exclusion regulator in lung adenocarcinoma. Journal for immunotherapy of cancer 2025 — PMC11865799.
- Identification and Validation of a Nine-Gene Amino Acid Metabolism-Related Risk Signature in HCC. Frontiers in cell and developmental biology 2021 — PMC8452960.
- PubMed PMID:9405390 — UniProt-cited evidence.
- PubMed PMID:9597550 — UniProt-cited evidence.
- PubMed PMID:11588157 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for B4GALT2 (Beta-1,4-galactosyltransferase 2). Expected localisation Golgi stack membrane expected; tissue compartment unverified (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A10774T2). Validated IHC antibody: A10774T2 at 1:100-1:300 (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 cervical carcinoma tissue; fixative not specified (datasheet A10774T2); 2. Fixation: Image fixative and duration unreported (datasheet A10774T2); 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 A10774T2); 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-B4GALT2, 1:100-1:300 (datasheet A10774T2); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining may confound interpretation (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope maps unknown; cytosolic versus lumenal location matters (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt O60909, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the Golgi apparatus and vesicles., Datasheet A10774T2, PMC11865799. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/B4GALT2/immunohistochemistry-ihc-guide.