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CHST12 / Carbohydrate sulfotransferase 12 · IHC design guide
Design Immunohistochemistry for CHST12
Plan CHST12 IHC-P with adrenal glandular cells as a high-staining reference and parathyroid glandular cells as a not-detected comparator (HPA tissue IHC). This guide covers fixation, the catalog antibody’s 1:25 starting dilution (datasheet A10456-1), and interpretation of cytoplasmic and nuclear tissue staining (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 | Tissue: cytoplasmic/nuclear (HPA tissue IHC); protein: Golgi membrane (UniProt) | |
| Staining pattern | Cytoplasmic and nuclear staining in most tissues (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 consistent fixation (selected-SKU IHC image A10456-1) | |
| Caveat | Antibody staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Widely expressed; no regulator specified (UniProt) | |
| Isoform / epitope | No isoforms annotated; N-tail cytoplasmic, bulk lumenal (UniProt) |
Section 1
Recommended CHST12 IHC & IF Protocols
The catalog antibody’s IHC protocol is accompanied by one published CHST12 protocol for glioblastoma and adjacent tissues (PMC8806823: Immunohistochemistry).
| 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 citrate pH 6 heat retrieval (datasheet: A10456-1); try the published EDTA microwave retrieval if staining needs optimization (PMC8806823: Immunohistochemistry).
Section 2
What Is the Expected CHST12 Staining Pattern?
CHST12 is a Golgi membrane protein with residues 1–5 cytoplasmic, residues 6–26 spanning the membrane, and residues 27–414 lumenal (UniProt Q9NRB3 topology). In paraffin sections, look for staining in glandular cells of adrenal gland and gallbladder and in skin melanocytes (HPA: High). HPA also reports cytoplasmic and nuclear staining in most tissues; its tissue IHC reliability is Approved, with medium consistency between staining and RNA data (HPA tissue IHC).
| Granular or compact perinuclear staining appears in adrenal or gallbladder glandular cells, or skin melanocytes. | This is compatible with Golgi localization (UniProt Q9NRB3; HPA ICC-IF: Golgi approved) in cell types with High tissue IHC staining (HPA tissue IHC). Judge the cell type and subcellular pattern separately: paraffin-section staining alone does not prove that every positive structure is Golgi. |
| Predominantly nuclear staining appears without a convincing cytoplasmic or perinuclear component. | Treat this as unresolved, rather than automatically calling it specific or artifactual. HPA reports cytoplasmic and nuclear staining in most tissues (HPA tissue IHC), while UniProt places CHST12 in the Golgi membrane and HPA ICC-IF assigns its main location to Golgi (UniProt Q9NRB3; HPA ICC-IF). Compare the pattern with a known-positive section and controls. |
| The strongest stain is in a cell population other than the expected positive population. | Check cell identity and the matched negative control before assigning CHST12. For example, HPA reports High staining in skin melanocytes, but does not thereby establish High staining in every skin cell (HPA tissue IHC). Unexpected staining may reflect cross-reactivity or endogenous chromogen-producing activity (general IHC practice); neither cause follows from the image alone. |
| Color spreads across tissue, stroma, or blank areas and obscures cell boundaries. | A diffuse deposit cannot be scored as the expected cellular pattern. HPA describes cytoplasmic and nuclear expression in most tissues, while its ICC-IF localization is Golgi (HPA tissue IHC; HPA ICC-IF). Review the negative control, blocking, washes, and detection chemistry for nonspecific or endogenous signal (general IHC practice). |
| A putative positive section has no detectable stain in its expected cells. | First confirm the sampled cell type: adrenal and gallbladder glandular cells and skin melanocytes are reported High by HPA (HPA tissue IHC). An absent signal could reflect assay performance or specimen variation; HPA staining levels are reference observations, not guarantees for every section. Check controls and the antibody's IHC instructions before interpreting absence biologically. |
💡Expected CHST12 appearanceA convincing positive shows cellular staining in adrenal or gallbladder glandular cells or skin melanocytes (HPA: High), preferably with a perinuclear Golgi-compatible component (UniProt Q9NRB3; HPA ICC-IF: Golgi); widespread structureless color is a suspect positive (general IHC practice).
| Membrane orientation and epitope location | CHST12 has one membrane-spanning segment at residues 6–26, a short cytoplasmic segment at 1–5, and a lumenal region at 27–414 (UniProt Q9NRB3 topology). Epitope location could matter when selecting or interpreting an antibody, but the supplied record gives no antibody epitope, so it cannot predict antigen-retrieval conditions. |
| Glycosylation | Four glycosylation sites are annotated at residues 134, 209, 280, and 370 (UniProt Q9NRB3). This identifies a potential consideration if an epitope is later mapped near those sites; the record does not show an effect on this antibody's IHC staining or justify a retrieval change. |
| Tissue reference and validation scope | HPA rates tissue IHC Approved with medium consistency between antibody staining and RNA expression; antibody HPA041680 is IHC Approved (HPA tissue IHC; HPA antibody record). Use the reported cell-level staining to choose comparison sections. Approval supports use as a reference, but does not make every stained compartment independently confirmed. |
| IF/ICC: what pattern should I expect? | HPA assigns an approved Golgi apparatus location in ICC-IF and lists images from A-431, U-251MG, and U2OS (HPA ICC-IF). This supports a Golgi-localization expectation for IF/ICC; it does not establish that a paraffin-section chromogenic image will resolve the same fine structure. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in adrenal or gallbladder glandular cells. | The section may lack suitable target cells, or the IHC run may have failed; both tissues have High staining in those cells in HPA's reference images (HPA tissue IHC). | Verify cell identity and section quality, inspect the run control, then follow the IHC-validated antibody's documented staining conditions. Do not infer CHST12 absence from a failed control (general IHC practice). |
| Only broad nuclear staining is visible. | Its meaning is ambiguous: HPA reports nuclear as well as cytoplasmic tissue staining, whereas UniProt and HPA ICC-IF support Golgi localization (HPA tissue IHC; UniProt Q9NRB3; HPA ICC-IF). | Compare a known-positive section, assess the negative control, and record nuclear and cytoplasmic scores separately. Avoid calling nuclear signal either definitive CHST12 or definitive artifact from this evidence alone. |
| Strong stain appears in expected-negative cells. | HPA reports no detection in cerebellar granular-layer cells, hippocampal glial cells, and parathyroid glandular cells (HPA tissue IHC). A conflicting result may be biological variation, cross-reactivity, or assay background. | Confirm the exact cell population and examine negative controls before scoring. Compare with a High-staining reference cell population in the same run (HPA tissue IHC; general IHC practice). |
| The entire section has diffuse chromogen. | Nonspecific binding, insufficient washing, or endogenous detection activity can produce broad color (general IHC practice). A diffuse field cannot establish the cell-specific HPA pattern. | Review the detection-only control, blocking, washes, and endogenous-activity control appropriate to the chromogen system; repeat scoring only after cell boundaries and compartments can be distinguished (general IHC practice). |
| Staining is patchy within a supposedly uniform positive tissue. | A tissue label does not define every cell's staining: HPA's High calls specify glandular cells or melanocytes (HPA tissue IHC). Section quality and uneven reagent coverage can also produce patchiness (general IHC practice). | Score the named cell population, inspect morphology and section coverage, and compare another area or section before treating patchiness as a CHST12 distribution change. |
| Perinuclear staining is difficult to distinguish from general cytoplasmic stain. | HPA's paraffin-tissue profile includes cytoplasmic staining, while the finer Golgi assignment comes from ICC-IF and UniProt localization (HPA tissue IHC; HPA ICC-IF; UniProt Q9NRB3). | Report the compartment visible at the section's resolution, with its cell type and intensity. Use the HPA ICC-IF result as localization context, without requiring a sharply resolved Golgi pattern for every chromogenic section. |
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 adrenal gland first: its glandular cells should stain strongly (HPA: High in adrenal gland glandular cells). Use parathyroid glandular cells as the negative tissue (HPA: Not detected); on the adrenal slide, adjacent nonglandular areas should lack the specific glandular staining pattern, although HPA does not establish them as CHST12-negative cells (HPA: High in adrenal gland glandular cells).
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 no-primary (secondary-only) and irrelevant primary controls matched to the antibody’s host species, immunoglobulin class, and clonality; use CHST12 knockout tissue or a validated peptide-block control as a biological specificity check (standard IHC practice). Quench endogenous peroxidase and check for background from the biotinylated secondary used in the selected protocol (A10456-1 tissue-IHC caption: biotinylated secondary).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10456-1 tissue-IHC record does not provide a reliable fixative assignment (caption metadata: fixative unreported). Heat retrieval in citrate buffer at pH 6 is reported, but whether retrieval is essential has not been established (A10456-1 tissue-IHC caption: citrate retrieval). The supplied evidence supports paraffin-section IHC; whether frozen sections or IF are easier is unreported, while endogenous peroxidase and biotin-related background warrant checks in the selected chromogenic workflow (A10456-1 tissue-IHC caption: biotinylated secondary; standard IHC practice).
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, cell type and compartment before interpreting chromogenic signal.
How should I adjust retrieval when CHST12 staining is weak in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 (datasheet A10456-1). The selected CHST12 tissue image used this retrieval on paraffin sections, followed by 3% BSA for 30 minutes and primary antibody at 1:25 for 1 hour at 37°C (caption A10456-1). If staining remains weak, compare a second retrieval condition on adjacent sections while keeping antibody concentration and detection constant; treat that condition as an optimization, not a validated CHST12 setting. Check whether signal appears in the expected glandular cells and has a compact Golgi-like distribution before accepting a stronger stain (HPA: high in adrenal gland glandular cells; HPA subcellular: Golgi apparatus).
Could fixation explain weak or uneven CHST12 staining?
Target-specific CHST12 sensitivity to fixation is unknown from the supplied evidence; do not infer it from tissue staining patterns or protein topology. For paraffin IHC, compare sections fixed and processed under documented, consistent conditions, then keep heat-mediated citrate retrieval at pH 6 constant during the comparison (datasheet A10456-1). Uneven fixation can produce spatial staining differences in routine IHC, so assess section thickness, tissue preservation and staining across the full section before changing the antibody dilution. If a processing variable correlates with signal, confirm it on replicate sections and judge the resulting pattern against Golgi localization and the expected cell population (HPA subcellular: Golgi apparatus; HPA: high in adrenal gland glandular cells).
What CHST12 staining pattern should I expect in chromogenic IHC?
Prioritize a compact perinuclear cytoplasmic pattern because CHST12 is a Golgi membrane protein (UniProt Q9NRB3 subcellular; HPA subcellular: Golgi apparatus). Its single transmembrane segment spans residues 6–26, with residues 27–414 on the lumenal side (UniProt Q9NRB3 topology). HPA tissue IHC also reports cytoplasmic and nuclear expression in most tissues, so a nuclear signal deserves assessment rather than automatic acceptance as Golgi staining (HPA: tissue profile). Compare the signal with cell morphology and a no-primary control, and examine intact central tissue as well as edges. A diffuse extracellular deposit or uniformly stained nuclei cannot, by itself, establish the expected subcellular pattern (UniProt Q9NRB3 subcellular).
How should CHST12 topology and epitope uncertainty affect IHC troubleshooting?
CHST12 has no annotated isoforms in the supplied record, so isoform selection offers no supported explanation for discrepant staining (UniProt Q9NRB3 isoforms). The record places residues 1–5 in the cytoplasm and 27–414 in the Golgi lumen, separated by a transmembrane segment at 6–26 (UniProt Q9NRB3 topology). Four glycosylation sites are listed at residues 134, 209, 280 and 370, but the antibody epitope is not supplied (UniProt Q9NRB3 glycosylation; supplied antibody evidence). If retrieval changes signal, compare localization and controls before attributing the difference to glycosylation or epitope masking. Record the antibody's epitope information if it becomes available, then interpret accessibility in light of its mapped location.
How can IF help verify an ambiguous CHST12 IHC pattern?
Use IF as a separate validation experiment and multiplex CHST12 with a marker verified for the expected cell population, such as the glandular cells scored in adrenal gland IHC (HPA: high in adrenal gland glandular cells). Add a Golgi marker to assess compartment agreement, since CHST12 is assigned to the Golgi apparatus (UniProt Q9NRB3 subcellular; HPA subcellular: Golgi apparatus). Choose fluorophores after inspecting unstained tissue in each channel, and place the weaker signal in a channel with lower tissue autofluorescence. Because the antibody epitope is unspecified, compare permeabilisation conditions cautiously: a lumenal epitope would require access across intracellular membranes, whereas residues 1–5 face the cytoplasm (UniProt Q9NRB3 topology).
What should I change if CHST12 IHC has widespread background?
First inspect the no-primary control and identify whether background follows tissue edges, damaged areas or the detection reagents. The selected tissue image used a 3% BSA, 30-minute block and a 1:25 primary incubation for 1 hour at 37°C; these are image conditions, not a universal optimum (caption A10456-1). For chromogenic detection, check the peroxidase block, wash stringency and development time while changing one variable at a time. If using a biotin-based secondary like the selected image, assess endogenous biotin with a matched control (caption A10456-1: biotinylated secondary). Retain conditions that resolve discrete cell staining consistent with Golgi localization (HPA subcellular: Golgi apparatus).
How should I quantify CHST12 positivity across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scoring compartment and eligible cells before reading the slides: CHST12 is assigned to the Golgi, while HPA tissue IHC describes broader cytoplasmic and nuclear staining (UniProt Q9NRB3 subcellular; HPA: tissue profile). For a specified cell population, report % positive cells and an H-score from 0 to 300 using consistent intensity thresholds; for sparse populations, report positive cells per mm² of viable tissue. Normalize counts to eligible cells or measured viable area, and exclude folds, edges and necrotic regions using prespecified rules. Score comparable anatomical regions and processing batches together, with blinded repeat reads where feasible. State the compartment counted so nuclear and Golgi-like signals are not silently combined.
When is a CHST12-positive IHC result convincing?
Look for reproducible staining in intact cells, with a compact Golgi-like component consistent with CHST12 localization (UniProt Q9NRB3 subcellular; HPA subcellular: Golgi apparatus). High staining in adrenal gland glandular cells or gallbladder glandular cells offers tissue context, while cerebellar granular-layer cells are reported as not detected; these are comparison patterns, not absolute controls (HPA: tissue IHC). Treat isolated nuclear-only, surface-wide or extracellular staining cautiously, particularly when it peaks at section edges or necrotic areas. Use a no-primary control and an appropriate detection-only control to check endogenous enzyme or detection-reagent signal. HPA assigns only medium consistency between antibody staining and RNA expression, so corroborate consequential conclusions independently (HPA: reliability description).
Boster reagents
Best CHST12 / Carbohydrate sulfotransferase 12 IHC Antibodies
The catalog provides real CHST12 IHC data from human small-intestine paraffin sections (A10456-1 image caption); it provides no IF figure (catalog: IF image alts).
Real IHC data
Anti-CHST12 Antibody (N-term)
Cat # A10456-1
A10456-1 is listed for human IHC-P (catalog: applications and reactivity). Its figure shows staining in human small-intestine paraffin sections fixed with formaldehyde, with citrate pH 6 heat retrieval (A10456-1 image caption).
Which to pick: For tissue IHC, choose A10456-1: this rabbit polyclonal antibody is listed for human IHC-P, and its own figure documents paraffin sections (catalog: host, clonality, applications and reactivity; A10456-1 image caption). No listed SKU has IF/ICC validation or an IF figure (catalog: applications and IF image alts). No listed SKU supports a cross-species choice because A10456-1 lists human reactivity only (catalog: reactivity).
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 Tissue: cytoplasmic/nuclear (HPA tissue IHC); protein: Golgi membrane (UniProt). 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 consistent fixation (selected-SKU IHC image A10456-1); Caveat: Antibody staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: No isoforms annotated; N-tail cytoplasmic, 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.