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CHST12 / Carbohydrate sulfotransferase 12 · IHC design guide
Design Immunohistochemistry for CHST12
Plan CHST12 IHC-P using the catalog antibody at 1:25 on formaldehyde-fixed paraffin sections (datasheet A10456-1). Assess cytoplasmic and nuclear staining, with high staining reported in adrenal and gallbladder glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CHST12 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 and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Most tissues: cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A10456-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Cerebellum+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Use formaldehyde-fixed paraffin sections and keep fixation consistent (selected-SKU IHC image A10456-1) | |
| Caveat | Nuclear staining differs from the Golgi localization (HPA tissue IHC; UniProt) | |
| Regulation | Widely expressed; no regulator specified (UniProt) | |
| Isoform / epitope | No isoforms; N-terminus cytosolic, bulk lumenal (UniProt) |
Section 1
Recommended CHST12 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A10456-1) with the published CHST12 protocol for GBM and adjacent tissues (PMC8806823).
| Sample | Formaldehyde-fixed, paraffin-embedded Human small intestine tissue (datasheet A10456-1) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A10456-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet A10456-1); 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-CHST12, 1:25 (datasheet A10456-1) |
| Primary incubation | 1 hours at 37°C (datasheet A10456-1) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CHST12-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet A10456-1); the GBM study used microwave EDTA retrieval (PMC8806823).
Section 2
What Is the Expected CHST12 Staining Pattern?
CHST12 is a Golgi membrane enzyme with a cytoplasmic N terminus and lumenal catalytic region (UniProt Q9NRB3 topology). In paraffin IHC, expect staining in glandular cells, melanocytes and other cell types reported by HPA, with intensity varying by tissue (HPA tissue IHC). HPA describes cytoplasmic and nuclear staining in most tissues; its IHC reliability is Approved, with medium consistency between staining and RNA data (HPA tissue IHC).
| Compact, perinuclear cytoplasmic staining in glandular cells, with stronger staining in adrenal gland or gallbladder. | A Golgi-associated pattern fits CHST12 localization (UniProt Q9NRB3; HPA subcellular ICC-IF). High staining in those glandular cells is reported, but their IHC images do not establish Golgi resolution (HPA tissue IHC). |
| Predominantly nuclear staining, with little convincing cytoplasmic signal. | Treat the compartment assignment as uncertain: Golgi is the approved subcellular location, while HPA also reports nuclear tissue IHC staining (HPA subcellular ICC-IF; HPA tissue IHC). Compare controls before calling nuclear-only signal CHST12 (general IHC practice). |
| Strong staining in a cell population that the selected HPA tissue entry reports as not detected. | Possible cross-reactivity or endogenous detection activity warrants checking, rather than an automatic positive call (general IHC practice). HPA's not-detected calls apply to specified cells, such as parathyroid glandular cells, not every cell in the tissue (HPA tissue IHC). |
| Diffuse staining across cells, stroma and spaces, without a discernible cellular pattern. | This does not provide convincing Golgi-associated localization (UniProt Q9NRB3; HPA subcellular ICC-IF). Review no-primary and detection controls for nonspecific background or endogenous detection activity (general IHC practice). |
| No staining in adrenal glandular cells or skin melanocytes. | These are reported High in HPA IHC, so an absent result raises concern about assay performance or specimen interpretation; it does not establish biological absence (HPA tissue IHC; general IHC practice). |
💡Expected CHST12 appearanceCall a positive when cellular staining includes a plausible perinuclear cytoplasmic component in the appropriate cells, especially High adrenal or gallbladder glandular cells or skin melanocytes (UniProt Q9NRB3; HPA subcellular ICC-IF; HPA tissue IHC); diffuse tissue-wide staining or an isolated nuclear signal needs control-based review (general IHC practice).
| Tissue and cell selection | HPA reports High glandular staining in adrenal gland and gallbladder, High melanocyte staining in skin, and Medium staining in several other listed populations (HPA tissue IHC). Low tissue RNA specificity and wide expression make a single tissue an incomplete specificity test (HPA tissue IHC; UniProt Q9NRB3). |
| Subcellular interpretation | CHST12 has a membrane span at residues 6–26, with residues 1–5 cytoplasmic and 27–414 lumenal (UniProt Q9NRB3 topology). Its approved ICC-IF location is Golgi, while the broader tissue IHC profile includes cytoplasmic and nuclear staining (HPA subcellular ICC-IF; HPA tissue IHC). |
| Antibody and epitope information | HPA041680 is Approved for IHC and ICC, but the supplied record gives no epitope position or target-specific retrieval response (HPA antibodies; supplied evidence scope). Select and document retrieval conditions using the antibody's IHC instructions and run appropriate controls (general IHC practice). |
| Protein annotations | UniProt lists one chain spanning residues 1–414, four glycosylation sites, and no annotated isoforms, signal peptide or propeptide (UniProt Q9NRB3). These annotations do not predict whether paraffin fixation or retrieval will change staining (supplied evidence scope). |
| IF/ICC question: what location should a separate IF guide assess? | The approved ICC-IF location is Golgi; HPA lists images from A-431, U-251MG and U2OS (HPA subcellular ICC-IF). The tissue IHC profile should not be used as an IF protocol or as proof of nuclear ICC-IF localization (HPA tissue IHC; HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| High HPA-positive cells give no chromogenic signal. | Possible assay failure, weak detection or mistaken cell identification; the HPA result alone cannot identify which (HPA tissue IHC; general IHC practice). | Confirm the cell population and compare a same-run positive control; review the antibody's documented IHC dilution, retrieval and detection steps (general IHC practice). |
| Staining is nuclear-dominant, with no plausible cytoplasmic component. | The result is discordant with the approved Golgi location, although nuclear tissue IHC staining is reported (HPA subcellular ICC-IF; HPA tissue IHC). | Compare positive and no-primary controls, examine cellular morphology, and score nuclear and cytoplasmic components separately (general IHC practice). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible; HPA's cell-level calls provide a comparison, not proof of assay specificity (HPA tissue IHC; general IHC practice). | Check the exact cell type against its HPA entry and inspect no-primary and detection controls before assigning CHST12 positivity (HPA tissue IHC; general IHC practice). |
| Diffuse background obscures cellular boundaries. | Nonspecific antibody or chromogen deposition, or endogenous detection activity, can obscure localization (general IHC practice). | Review blocking, washes, antibody concentration and detection controls; reassess only areas where cellular staining can be distinguished (general IHC practice). |
| Only faint staining appears in a reported High population. | The selected section or run may differ from the HPA example; High is an observed HPA category, not a guaranteed intensity in every preparation (HPA tissue IHC; general IHC practice). | Verify morphology, a same-run positive control and the catalog antibody's IHC instructions before interpreting weak signal as low expression (general IHC practice). |
| A presumed negative area shows scattered positive cells. | A not-detected HPA call may refer to one named cell population, such as hippocampal glial cells or cerebellar granular-layer cells (HPA tissue IHC). | Identify the stained cells and compare the matching HPA cell-level entry; do not score the whole tissue as a uniform negative control (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CHST12 staining
A10456-1 · IHCA10456-1 staining CHST12 in Human small intestine tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136213-CHST12/tissue/cerebral+cortex#imid_11067215
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000136213-CHST12/tissue/adrenal+gland#imid_11067175
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136213-CHST12/tissue/colon#imid_11085629
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136213-CHST12/tissue/liver#imid_11085630
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041680; green = CHST12, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041680; green = CHST12, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CHST12 IHC & IF
🧪Run gallbladder first and expect staining in its glandular cells (HPA: High in gallbladder glandular cells). Use parathyroid glandular cells as the negative tissue (HPA: Not detected); on the gallbladder slide, non-glandular cells can serve as internal background comparators only where they remain unstained.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Cerebellum (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHST12 in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and monoclonal or polyclonal format; and CHST12-knockout material as a biological negative (standard IHC practice). Quench endogenous peroxidase and assess endogenous biotin when using the caption’s biotinylated secondary antibody (selected-SKU IHC caption: biotinylated secondary).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU record lists the fixative as unreported (selected-SKU IHC caption). The caption uses heat retrieval in citrate buffer at pH 6, but does not establish that CHST12 staining requires it (selected-SKU IHC caption). ICC-IF shows Golgi localization, while frozen-section performance and whether IF is easier are unreported (HPA: approved Golgi apparatus localization); check for endogenous biotin signal with the caption’s biotinylated detection method (selected-SKU IHC caption).
HPA tissue IHC evidence for CHST12
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Skin | Melanocytes | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced CHST12 IHC Tips
Troubleshoot CHST12 staining in paraffin sections by checking retrieval, compartment, controls and cell-specific scoring before interpreting chromogenic signal.
What retrieval should I start with if CHST12 staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 (datasheet A10456-1). The selected small-intestine IHC image used that retrieval with primary antibody at 1:25 for 1 hour at 37°C (caption A10456-1). If staining is weak, compare a modestly longer heating interval on matched sections while keeping the buffer, detection chemistry and antibody dilution constant (standard IHC practice). Include a no-primary control and assess tissue integrity, because stronger retrieval can increase nonspecific staining or damage morphology (standard IHC practice). Score any improvement in cells and compartments consistent with Golgi localisation, rather than by overall darkness alone (UniProt Q9NRB3 localisation; HPA subcellular: Golgi apparatus).
How should I troubleshoot variable CHST12 staining across differently fixed blocks?
CHST12-specific fixation sensitivity is unknown from the supplied evidence, so treat differences among blocks as an experimental variable rather than an established property of the target. For paraffin IHC, compare sections processed with documented fixative type and fixation duration, using the same retrieval, antibody incubation and chromogen development in one run (standard IHC practice). Start with citrate at pH 6 and keep the tested 1:25 primary dilution as a reference condition (datasheet A10456-1; caption A10456-1). Review morphology and matched negative controls before attributing weaker staining to antigen loss, and avoid comparing absolute intensity across differently processed blocks without controls (standard IHC practice).
Does diffuse cytoplasmic or nuclear CHST12 staining fit the expected localisation?
CHST12 is annotated as a Golgi membrane protein, with a transmembrane segment at residues 6–26 and a lumenal region at 27–414 (UniProt Q9NRB3 topology and localisation). Look for a compact perinuclear cytoplasmic pattern when morphology permits subcellular assessment; chromogenic paraffin sections may not resolve individual Golgi structures (HPA subcellular: Golgi apparatus; standard IHC practice). Tissue IHC reports cytoplasmic and nuclear staining in most tissues, whereas the approved subcellular location is Golgi (HPA tissue IHC profile; HPA subcellular). Treat isolated nuclear signal cautiously and compare it with no-primary staining, nearby cell types and an independently validated Golgi marker before assigning it to CHST12 (standard IHC practice).
Could epitope position explain discordant CHST12 staining patterns?
The supplied record lists 0 annotated isoforms, so an isoform explanation for discordant staining is unsupported here (UniProt Q9NRB3 isoforms). CHST12 has a short cytoplasmic segment at residues 1–5, a membrane span at 6–26 and a lumenal region at 27–414 (UniProt Q9NRB3 topology). Four glycosylation sites are annotated at residues 134, 209, 280 and 370, but their effects on this antibody’s binding are unknown (UniProt Q9NRB3 glycosylation). Obtain the antibody’s immunogen coordinates, if available, before testing whether retrieval or accessibility explains a mismatch; compare matched sections with a second, independently mapped antibody if interpretation remains uncertain (standard IHC practice).
How can IF help assess an ambiguous CHST12 IHC pattern?
Use IF as a separate localisation check, since CHST12 has an approved Golgi location and IF/ICC images are listed for A-431, U-251MG and U2OS cells (HPA subcellular). Multiplex CHST12 with a validated marker for the cell type identified by morphology, then assess whether signal occupies the expected cells as well as the expected compartment (standard IF practice; HPA subcellular: Golgi apparatus). Choose a fluorophore channel after measuring tissue autofluorescence in an unstained control, and verify spectral separation with single-stain controls (standard IF practice). Permeabilise according to the mapped epitope: access to the lumenal residues 27–414 requires membrane access, while the cytoplasmic tail occupies residues 1–5; the catalog antibody’s epitope is unspecified (UniProt Q9NRB3 topology).
What should I check when CHST12 DAB staining is widespread?
Check a no-primary section and a secondary-only control to separate tissue or detection background from antibody-dependent staining (standard IHC practice). The selected image used 3% BSA for 0.5 hour and a biotinylated secondary antibody, so test a biotin-blocking step if the same detection system gives widespread signal (caption A10456-1; standard IHC practice). Include a peroxidase block before DAB development and compare development times across matched sections (standard chromogenic IHC practice). Because CHST12 is Golgi-localised, diffuse extracellular deposit, uniform nuclear colour or staining concentrated at section edges needs control-supported verification before interpretation (HPA subcellular: Golgi apparatus; standard IHC practice).
How should I quantify CHST12 staining without overcalling background? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; CHST12 is Golgi-localised, while tissue IHC also describes broad cytoplasmic and nuclear staining (HPA subcellular; HPA tissue IHC profile). For interpretable cells, record percent positive and intensity as an H-score, using a prespecified threshold established with no-primary and reference sections (standard IHC scoring practice). Alternatively, count positive cells per mm² in a consistent tissue compartment, excluding folds, necrosis and section edges (standard IHC scoring practice). Normalise to the number of eligible cells or the measured viable area, and compare only sections processed and developed together (standard IHC scoring practice).
When is a CHST12-positive IHC field convincing?
A convincing field shows reproducible staining in identifiable cells with a plausible perinuclear pattern, supported by controls and the approved Golgi localisation (HPA subcellular: Golgi apparatus; standard IHC practice). Glandular cells in adrenal gland and gallbladder have reported high staining, whereas cerebellar granular-layer cells and parathyroid glandular cells are reported as not detected (HPA tissue IHC); use such references only when those tissues are actually in the study. Nuclear-only staining, unexpected cell populations, edge accentuation and necrotic deposits warrant review against matched controls before assignment to CHST12 (HPA tissue IHC profile; standard IHC practice). Check no-primary and peroxidase-block controls to distinguish antibody-dependent signal from endogenous enzyme activity or DAB artefact (standard chromogenic IHC practice).
Boster reagents
Best CHST12 / Carbohydrate sulfotransferase 12 IHC Antibodies
A10456-1 has real IHC data from human small intestine paraffin sections (catalog image caption). No IF data are supplied (catalog IF images).
Real IHC data
Anti-CHST12 Antibody (N-term)
Cat # A10456-1
A10456-1 is listed for human IHC-P (catalog applications/reactivity). Its IHC image shows CHST12 staining in human small intestine paraffin sections (catalog image caption).
Which to pick: Choose A10456-1 for human tissue IHC-P: it is a rabbit polyclonal antibody with human reactivity and a paraffin-section IHC image (catalog antibody details; catalog image caption). The caption describes the sections as paraformaldehyde-fixed and also reports formaldehyde fixation (catalog image caption). No listed SKU has IF/ICC or nonhuman validation, so the payload supports no choice for those uses (catalog applications/reactivity; catalog IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NRB3 (CHSTC_HUMAN, Carbohydrate sulfotransferase 12).
- Human Protein Atlas. CHST12 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CHST12 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. CHST12 antibody validation summary (1 antibodies).
- Knockdown of carbohydrate sulfotransferase 12 decreases the proliferation and mobility of glioblastoma cells via the WNT/β-catenin pathway. Bioengineered 2021 — PMC8806823.
- PubMed PMID:10781601 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CHST12 (Carbohydrate sulfotransferase 12). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A10456-1). Validated IHC antibody: A10456-1 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Cerebellum. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded Human small intestine tissue (datasheet A10456-1); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A10456-1); 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 retrieval: Citrate pH 6 (datasheet A10456-1); 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-CHST12, 1:25 (datasheet A10456-1); 9. Primary incubation: 1 hours at 37°C (datasheet A10456-1). Watch for: Fixation: Use formaldehyde-fixed paraffin sections and keep fixation consistent (selected-SKU IHC image A10456-1); Caveat: Nuclear staining differs from the Golgi localization (HPA tissue IHC; UniProt); Isoform / epitope: No isoforms; N-terminus cytosolic, bulk lumenal (UniProt); Regulation: Widely expressed; no regulator specified (UniProt). Evidence: UniProt Q9NRB3, HPA tissue IHC · Approved, HPA ICC-IF · Golgi apparatus., Datasheet A10456-1, PMC8806823. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CHST12/immunohistochemistry-ihc-guide.