This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CSGALNACT2 / Chondroitin sulfate N-acetylgalactosaminyltransferase 2 · IHC design guide
Design Immunohistochemistry for CSGALNACT2
Plan paraffin-section IHC for CSGALNACT2 using granular cytoplasmic tissue staining as the observed pattern and Golgi localization as molecular context (HPA tissue IHC; UniProt). Use adrenal gland glandular cells as a high-staining control (HPA tissue IHC), and optimize the catalog antibody within 1:100–1:300 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CSGALNACT2 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 observed; Golgi membrane expected (HPA tissue IHC; UniProt) | |
| Staining pattern | Granular cytoplasmic staining in most tissues (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 | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10939) | |
| Caveat | Staining shows low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Specific expression regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope map unknown; cytosolic versus lumenal side matters (UniProt) |
Section 1
Recommended CSGALNACT2 IHC & IF Protocols
Use the catalog antibody’s IHC-P protocol (datasheet) alongside the published ovarian cancer tissue protocol (PMC11219422).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10939) |
| Fixation | Image fixative and duration unreported (datasheet A10939); 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-CSGALNACT2, 1:100-1:300 (datasheet A10939) |
| 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 | CSGALNACT2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with the page’s citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); compare the published pressure-cooker condition if needed (PMC11219422).
Section 2
What Is the Expected CSGALNACT2 Staining Pattern?
CSGALNACT2 is a Golgi stack membrane protein with a short cytoplasmic segment and a large lumenal region (UniProt Q8N6G5 topology). In paraffin sections, expect granular cytoplasmic staining in tissue cells, including strong staining in several epithelial, glandular, glial, and endothelial populations (HPA tissue IHC). Interpret intensity cautiously: HPA rates the tissue staining Approved but reports low consistency with RNA expression (HPA tissue IHC).
| Granular cytoplasmic staining in the relevant cells, with little nuclear or diffuse background. | This matches the broad tissue pattern reported by HPA (HPA tissue IHC). A Golgi-associated pattern is biologically plausible for this membrane enzyme (UniProt Q8N6G5 localisation and topology). In a paraffin section, granules may not resolve into a distinct Golgi stack (general IHC practice). |
| Predominantly nuclear staining, or a crisp cell-surface outline without convincing cytoplasmic granules. | Neither pattern fits the reported Golgi membrane localisation or granular tissue staining (UniProt Q8N6G5 localisation; HPA tissue IHC). Treat it as suspect and assess controls before assigning it to CSGALNACT2; appearance alone cannot identify the source of an artefact (general IHC practice). |
| Strong staining appears chiefly in an unexpected cell population while the expected population is weak. | Consider cross-reactivity or endogenous chromogenic activity, depending on the detection system (general IHC practice). HPA documents high staining in selected cell types, but its list is not an exhaustive map: UniProt describes tissue expression as ubiquitous (HPA tissue IHC; UniProt Q8N6G5 tissue specificity). |
| Broad, featureless colour covers cells and surrounding tissue. | Diffuse background makes the granular cytoplasmic pattern difficult to judge (HPA tissue IHC). Non-specific antibody binding, inadequate washing, or endogenous detection activity are possible workflow causes; their presence must be checked with appropriate controls (general IHC practice). |
| No convincing stain appears in a section containing a cell population reported as strongly positive. | A failed stain is possible, but the HPA level is an observation, not a guarantee for every specimen (HPA tissue IHC). Check the run and tissue integrity first; HPA's reported low staining–RNA consistency limits conclusions drawn from one negative section (HPA tissue IHC; general IHC practice). |
💡Expected CSGALNACT2 appearanceCall a result compatible when discrete granular cytoplasmic staining is visible in relevant cells, potentially strong in HPA-listed high populations; predominantly nuclear, surface-only, or diffuse colour is suspect (HPA tissue IHC; UniProt Q8N6G5 localisation; general IHC practice).
| Compartment and epitope access | The protein has one membrane span at residues 12–32; residues 33–542 are lumenal (UniProt Q8N6G5 topology). The antibody epitope is unspecified here, so topology alone cannot determine an antigen retrieval condition or explain a failed paraffin stain. |
| Tissue choice | HPA reports high staining in bronchial respiratory epithelium and glandular cells in adrenal gland, appendix, and endometrium; it also reports high staining in selected glial and endothelial populations (HPA tissue IHC). Use the stated cell population, not the tissue name alone, when reading a section. |
| Evidence strength | HPA rates the listed antibody's IHC staining Approved and reports low consistency between staining and RNA expression (HPA antibody validation; HPA tissue IHC). A matching pattern supports interpretation but does not establish specificity in an individual run. |
| IF/ICC Q: Should the same compartment be expected? | HPA's approved ICC-IF location is endoplasmic reticulum, whereas UniProt assigns CSGALNACT2 to the Golgi stack membrane (HPA subcellular; UniProt Q8N6G5 localisation). Record the discrepancy when comparing images; the supplied evidence does not resolve it or justify an IF protocol here. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected granular cytoplasmic signal is absent throughout the run. | A run-level staining or detection failure is possible (general IHC practice). | Review the run control, reagent sequence, antigen retrieval record, and chromogen development before interpreting tissue as negative (general IHC practice). No target-specific fixation sensitivity is reported in the supplied UniProt or HPA evidence. |
| An HPA-listed high cell population is weak in one section. | The reported intensity may vary between specimens, or section quality may limit interpretation (HPA tissue IHC; general IHC practice). | Confirm that the named cell population is present and preserved; compare an independently stained section and run control (general IHC practice). Avoid treating HPA's high label as a required intensity for every specimen (HPA tissue IHC). |
| Nuclei dominate the staining. | This conflicts with Golgi membrane localisation and HPA's granular cytoplasmic tissue profile (UniProt Q8N6G5 localisation; HPA tissue IHC). | Inspect a negative reagent control and repeat the stain if needed; score nuclear-only colour as unconfirmed CSGALNACT2 (general IHC practice). |
| Unexpected cells stain strongly in a chromogenic section. | Cross-reactivity or endogenous activity in the chosen detection system is possible (general IHC practice). | Compare reagent controls and the distribution of cytoplasmic granules; check an HPA-described population in the same or a companion section (HPA tissue IHC; general IHC practice). Do not exclude a cell solely because HPA did not list it (UniProt Q8N6G5 tissue specificity). |
| Diffuse colour obscures cell boundaries. | Non-specific binding, residual detection reagents, or endogenous chromogenic activity may contribute (general IHC practice). | Check negative controls and washing and blocking steps; assess whether discrete cytoplasmic granules remain visible before scoring (general IHC practice; HPA tissue IHC). |
| An IF image suggests ER while the IHC section appears Golgi-associated. | The supplied location annotations differ by source and application (HPA subcellular; UniProt Q8N6G5 localisation). | Report the observed compartment and method separately. Do not use the IF annotation alone to relabel a granular paraffin-section result or to establish antibody specificity (HPA subcellular; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing CSGALNACT2 staining
A10939 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using CSGALNACT2 Antibody. The picture on the right is blo
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169826-CSGALNACT2/tissue/cerebral+cortex#imid_7876344
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169826-CSGALNACT2/tissue/colon#imid_7872886
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169826-CSGALNACT2/tissue/liver#imid_7872895
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169826-CSGALNACT2/tissue/kidney#imid_7872896
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031540; green = CSGALNACT2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA031540; green = CSGALNACT2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CSGALNACT2 IHC & IF
🧪Run appendix first and assess staining in its glandular cells (HPA: High in appendix glandular cells). HPA detects CSGALNACT2 in all 45 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and expect any unstained cells on the positive slide to show background chromogen only without treating them as confirmed target-negative cells (HPA: detected in all 45 scored tissues; UniProt: ubiquitous).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: CSGALNACT2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CSGALNACT2 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Run a no-primary (secondary-only) control and an isotype control matched to the primary antibody’s host species and clonality; use CSGALNACT2 knockout material if available or a matched peptide-block control as a biological specificity check (selected-SKU tissue-IHC caption: peptide block). For chromogenic appendix IHC, quench endogenous peroxidase, check background in inflammatory cells, and block endogenous biotin if using avidin-biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative is unreported in the A10939 paraffin-section caption (selected-SKU tissue-IHC caption). Retrieval dependence is unreported, so optimize antigen retrieval empirically; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU tissue-IHC caption; HPA: ICC-IF images in A-431, U-251MG and U2OS). In appendix, inflammatory-cell endogenous peroxidase can mimic chromogenic signal (standard IHC practice).
HPA tissue IHC evidence for CSGALNACT2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CSGALNACT2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CSGALNACT2 IHC Tips
Troubleshoot CSGALNACT2 staining by checking retrieval, compartment, cell identity and controls before comparing IHC scores across sections.
How should I optimize retrieval when CSGALNACT2 staining is weak?
Start with citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Keep section thickness, cooling time and detection conditions constant while comparing retrieved sections with a no-retrieval control (standard IHC practice). If staining remains weak, test a more alkaline retrieval buffer only as a fallback, using matched sections and monitoring tissue damage alongside signal (standard IHC practice). Score improvement in viable cells with granular cytoplasmic staining, rather than accepting stronger diffuse staining alone (HPA tissue IHC: granular cytoplasmic expression; standard IHC practice).
Could fixation explain variable CSGALNACT2 staining between blocks?
Target-specific fixation sensitivity is unknown: the A10939 image shows paraffin-embedded human breast carcinoma, but its caption does not report a fixative (A10939 caption). For prospective IHC comparisons, record the fixative and processing interval, and prepare matched blocks under the same conditions (standard IHC practice). Cut sections at a consistent 4 µm thickness and process comparison blocks together so differences in section preparation do not confound staining (standard IHC practice). If archival blocks differ, interpret any intensity shift alongside tissue preservation and matched controls; do not attribute it specifically to CSGALNACT2 fixation sensitivity (standard IHC practice).
What staining pattern should count as correctly localised CSGALNACT2?
Evaluate granular cytoplasmic staining within identifiable cells, the tissue pattern reported for CSGALNACT2, and record its distribution separately from diffuse cytoplasmic color (HPA tissue IHC). A perinuclear punctate pattern is plausible because UniProt assigns the protein to Golgi stack membrane, while HPA's approved cell-image localisation is endoplasmic reticulum (UniProt Q8N6G5 subcellular; HPA subcellular). Those sources differ, so morphology alone cannot settle the precise organelle in a chromogenic section (UniProt Q8N6G5 subcellular; HPA subcellular). Document whether signal follows intact cells rather than luminal debris or tissue edges, using the counterstain to define cell boundaries (standard IHC practice).
Can this stain distinguish CSGALNACT2 isoforms or reveal an inaccessible epitope?
Do not assign staining to one isoform: UniProt lists 2 isoforms, while the supplied A10939 caption gives no antibody epitope or isoform specificity (UniProt Q8N6G5 isoforms; A10939 caption). Check the antibody's documented immunogen against both isoform sequences before making an isoform claim, and keep any unmapped result at the gene-product level (standard IHC practice). The protein has a short cytoplasmic segment at residues 1–11 and a lumenal region at 33–542, so epitope position matters for access (UniProt Q8N6G5 topology). Glycosylation at residues 41 and 333 warrants caution when interpreting epitope-dependent staining; the peptide-blocked A10939 image supports competition in that specimen, not isoform discrimination (UniProt Q8N6G5 glycosylation; A10939 caption).
How should I check CSGALNACT2 by multiplex IF?
For a secondary IF check, pair CSGALNACT2 with an epithelial marker when examining bronchus, where HPA reports high staining in respiratory epithelial cells (HPA tissue IHC: bronchus). Select spectrally separated dyes and consider a far-red channel for the weaker signal after inspecting unstained tissue for autofluorescence (standard IF practice). UniProt places residues 33–542 on the lumenal side of the Golgi membrane, so an epitope there requires access through intracellular membranes; the supplied caption does not map the antibody epitope (UniProt Q8N6G5 topology; A10939 caption). Compare gentle permeabilisation conditions and include single-color controls, then assess overlap without treating it as proof of organelle identity (standard IF practice).
How can I separate CSGALNACT2 signal from chromogenic background?
Inspect a no-primary control and block endogenous peroxidase before DAB development so native enzyme activity is less likely to be mistaken for antibody signal (standard chromogenic IHC practice). Compare controls on sections processed with the same retrieval, secondary reagent and development time, particularly where edge staining or deposits appear (standard IHC practice). The A10939 tissue image includes a panel blocked with synthesized peptide; loss of staining in that comparison supports peptide competition under its pictured conditions (A10939 caption). It does not establish that every stained cell is specific, so judge granular cytoplasmic signal against tissue morphology and matched controls (HPA tissue IHC; standard IHC practice).
How should I quantify CSGALNACT2 across heterogeneous IHC sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the viable cell population and compartment before scoring, because HPA reports granular cytoplasmic expression in most tissues and lists different highly stained cell types (HPA tissue IHC). For cell-based analysis, record percent positive cells and an H-score from 0–300, using the same intensity thresholds across the batch (standard IHC practice). If the target population is sparse, report positive cells per mm² of viable annotated tissue and the number of cells assessed (standard IHC practice). Normalize comparisons to the relevant cell count or viable tissue area, and exclude necrosis and blank spaces from the denominator (standard IHC practice).
When is an apparent CSGALNACT2-positive region convincing?
A convincing region contains intact, identifiable cells with granular cytoplasmic signal, consistent with the reported tissue pattern (HPA tissue IHC). Check cell identity against the sampled tissue: HPA reports high staining in bronchial respiratory epithelial cells and colon endothelial cells, so whole-section intensity can obscure which cells contribute (HPA tissue IHC). Scrutinize diffuse extracellular color, necrotic areas, section edges and staining retained in no-primary controls before calling a region positive (standard IHC practice). Treat the peptide-blocked A10939 image as supporting competition in that specimen, while weighing conclusions against HPA's stated low consistency between antibody staining and RNA expression (A10939 caption; HPA tissue IHC reliability).
Boster reagents
Best CSGALNACT2 / Chondroitin sulfate N-acetylgalactosaminyltransferase 2 IHC Antibodies
A10939 has a human breast carcinoma paraffin-section IHC image with peptide blocking (catalog 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-CsGalNAcT-2 Antibody
Cat # A10939
A10939 is the only rendering SKU; its IHC figure shows paraffin-embedded human breast carcinoma tissue alongside a peptide-blocked image (catalog image caption). A10939 lists IHC and IF applications and Human/Mouse reactivity (catalog applications/reactivity), with no IF image supplied (catalog IF image alts).
Which to pick: Choose A10939 for paraffin-section tissue IHC based on its human breast carcinoma figure (catalog image caption); the fixative is unreported (catalog image caption). For IF/ICC, A10939 lists IF but has no IF image or ICC application listed, so validate it for the intended preparation (catalog applications/IF image alts). For Human/Mouse work, A10939 is a rabbit antibody listing both species (catalog host/reactivity), though its IHC figure shows human tissue only (catalog image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N6G5 (CGAT2_HUMAN, Chondroitin sulfate N-acetylgalactosaminyltransferase 2).
- Human Protein Atlas. CSGALNACT2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CSGALNACT2 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. CSGALNACT2 antibody validation summary (1 antibodies).
- CSGALNACT2 restricts ovarian cancer migration and invasion by modulating MAPK/ERK pathway through DUSP1. Cellular oncology (Dordrecht, Netherlands) 2024 — PMC11219422.
- PubMed PMID:12446672 — UniProt-cited evidence.
- PubMed PMID:12433924 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CSGALNACT2 (Chondroitin sulfate N-acetylgalactosaminyltransferase 2). Expected localisation Granular cytoplasm observed; Golgi membrane expected (HPA tissue IHC; UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10939 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A10939); 2. Fixation: Image fixative and duration unreported (datasheet A10939); 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-CSGALNACT2, 1:100-1:300 (datasheet A10939); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10939); Caveat: Staining shows low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope map unknown; cytosolic versus lumenal side matters (UniProt); Regulation: Specific expression regulation is unreported (UniProt). Evidence: UniProt Q8N6G5, HPA tissue IHC · Approved, HPA ICC-IF · endoplasmic reticulum., Datasheet A10939, PMC11219422. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CSGALNACT2/immunohistochemistry-ihc-guide.