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ARSB / Arylsulfatase B · IHC design guide
Design Immunohistochemistry for ARSB
Plan ARSB IHC-P around granular cytoplasmic staining and high-staining kidney tubules, cardiomyocytes, and cervical glandular cells (HPA tissue IHC). The IHC-validated antibody A01609 has been tested in human tissue (datasheet); assess staining by cell type and include a negative control (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ARSB 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 (HPA tissue IHC); lysosome and cell surface expected (UniProt) | |
| Staining pattern | Cytoplasmic granules; high in kidney tubules and cardiomyocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Ovary |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Ovarian stroma may lack staining (HPA tissue IHC) | |
| Regulation | Expression regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; signal peptide aa 1–36 is removed (UniProt) |
Section 1
Recommended ARSB IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by two published ARSB chromogenic IHC protocols (PMC4125552; PMC3866634).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A01609); 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-ARSB, 1:100–1:500 starting range (standard) |
| 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 | ARSB-positive staining in glandular cells of cervix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression often with a granular pattern. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule).
Section 2
What Is the Expected ARSB Staining Pattern?
ARSB should appear chiefly as granular cytoplasmic staining in paraffin tissue sections, with strong staining reported in cervical glandular cells, cardiomyocytes, and kidney tubule cells (HPA: tissue IHC). UniProt places ARSB in lysosomes and at the cell surface and reports no transmembrane segment (UniProt P15848 topology). HPA rates its tissue IHC profile Approved, with medium consistency between staining and RNA data and external verification pending (HPA: tissue IHC).
| Granular cytoplasmic staining is clear in cervical glandular cells, cardiomyocytes, or kidney tubule cells. | This matches the reported High staining in those cells and the broader granular cytoplasmic tissue profile (HPA: tissue IHC). Compare the signal with adjacent cell types on the same section before calling the entire tissue uniformly positive (general IHC practice). |
| Nuclear staining dominates while cytoplasmic granules are absent. | A nuclear dominant pattern does not match HPA tissue IHC or UniProt localization (HPA: tissue IHC; UniProt P15848 subcellular location). Treat it as suspect staining and assess background, counterstain, and antibody specificity before assigning ARSB expression (general IHC practice). |
| Ovarian stroma stains as strongly as a known positive cell population. | HPA reports ARSB as Not detected in ovarian stroma cells, although that observation does not establish a negative result for every ovarian cell (HPA: tissue IHC). Check whether the signal follows tissue structure or resembles nonspecific antibody binding or detection activity (general IHC practice). |
| Broad, diffuse color covers cells and surrounding tissue without discernible cytoplasmic granules. | This obscures the reported granular pattern and makes localization unreliable (HPA: tissue IHC). Uneven blocking, excess antibody, or detection background are possible workflow causes; inspect a control without primary antibody before interpreting the color as ARSB (general IHC practice). |
| Cervical glandular cells, cardiomyocytes, or kidney tubule cells show no convincing staining. | These cell populations were reported as High, so an absent signal calls for a run check, though HPA's Approved rating still has medium RNA agreement and awaits external verification (HPA: tissue IHC). Examine tissue preservation, retrieval, and detection controls before concluding that ARSB is absent (general IHC practice). |
💡Expected ARSB appearanceCall an IHC result positive when distinct granular cytoplasmic staining is evident in the relevant cells, especially the High cervical glandular, cardiac muscle, or kidney tubule populations; isolated nuclear color or diffuse section wide haze is suspect (HPA: tissue IHC; UniProt P15848 subcellular location; general IHC practice).
| Choice of reference tissue and cell population | Use the named High populations as positive comparators; ovarian stroma cells were Not detected (HPA: tissue IHC). HPA also describes widespread expression, so a low staining area elsewhere is not an automatic negative control (HPA: tissue IHC). |
| Compartment interpretation across methods | UniProt lists lysosomes and cell surface, while HPA describes granular cytoplasm in tissue IHC (UniProt P15848 subcellular location; HPA: tissue IHC). HPA's approved ICC/IF localization is mainly Golgi; it comes from a different assay and should not replace the tissue IHC reading rule (HPA: subcellular ICC/IF). |
| Antibody validation scope | The listed antibodies have IHC status Approved; HPA037770 also has ICC status Approved, while no ICC status is supplied for HPA037771 (HPA: antibodies). Approved tissue IHC has medium consistency with RNA and is pending external verification; avoid treating a single matching stain as independent proof of identity (HPA: tissue IHC). |
| Protein maturation and sequence coverage | UniProt records a signal peptide at residues 1–36, a chain at 37–533, 6 glycosylation sites, and 2 isoforms (UniProt P15848 processing, glycosylation, isoforms). The supplied data do not identify antibody epitopes, so they cannot predict whether processing or an isoform changes this antibody's stain. |
| Retrieval and detection settings | No ARSB specific retrieval condition or fixation sensitivity is supplied. Optimize retrieval and antibody concentration against positive tissue and a control without primary antibody, and keep detection exposure consistent when comparing sections (general IHC practice). Do not infer an ARSB fixation effect from HPA staining levels or UniProt modifications. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue has little or no cytoplasmic signal. | A failed staining run or unsuitable assay settings are possible; HPA reports High staining in the named reference cells (HPA: tissue IHC; general IHC practice). | Check a positive reference section, reagent performance, retrieval settings, and detection steps in the same run before scoring the specimen negative (general IHC practice). |
| Color is mostly nuclear or otherwise lacks the expected granular cytoplasmic pattern. | The location conflicts with tissue IHC and UniProt localization; background or an unrelated target may be contributing (HPA: tissue IHC; UniProt P15848 subcellular location; general IHC practice). | Review the counterstain and a control without primary antibody, then repeat with adjusted antibody concentration if the pattern remains unclear (general IHC practice). |
| Ovarian stroma cells appear strongly positive. | This differs from HPA's Not detected observation for that cell population; nonspecific binding or detection activity is possible (HPA: tissue IHC; general IHC practice). | Verify that the stained cells are stromal cells, inspect the control without primary antibody, and compare with a known positive population (general IHC practice). |
| Diffuse color prevents cell by cell scoring. | Background can mask the granular cytoplasmic profile reported by HPA (HPA: tissue IHC; general IHC practice). | Review blocking, antibody concentration, washes, and chromogen development with the same tissue and controls; score only clearly localized signal (general IHC practice). |
| A weak area is proposed as the sole negative control. | ARSB has a ubiquitous cytoplasmic tissue profile and low tissue RNA specificity; HPA reports low staining in some cells rather than universal absence (HPA: tissue IHC). | Use cell level comparisons and a control without primary antibody; if using ovarian stroma, limit the negative expectation to its reported cell population (HPA: tissue IHC; general IHC practice). |
| IF/ICC shows a Golgi concentrated pattern instead of the tissue IHC granules: is that inconsistent? | HPA reports mainly Golgi localization in ICC/IF, while its tissue IHC profile is granular cytoplasm (HPA: subcellular ICC/IF; HPA: tissue IHC). | Interpret each assay against its own HPA observation; use the separate IF/ICC guide for that assay's design and controls (HPA: subcellular ICC/IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing ARSB staining
A01609 · IHCImmunohistochemistry of ARSB in human lung tissue with ARSB antibody at 2.5 μg/mL.
A01609 · IFImmunofluorescence of ARSB in human lung tissue with ARSB antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000113273-ARSB/tissue/cerebral+cortex#imid_9286424
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000113273-ARSB/tissue/colon#imid_9279654
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000113273-ARSB/tissue/liver#imid_9279655
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000113273-ARSB/tissue/kidney#imid_9279656
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037770; green = ARSB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037770; green = ARSB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ARSB IHC & IF
🧪Run kidney first: cells in tubules should stain (HPA: High in kidney tubules). Use ovary as the negative tissue, focusing on ovarian stroma cells (HPA: Not detected in ovarian stroma); cells outside the stained tubules on the kidney slide should remain at background where they show no specific signal (standard IHC practice).
Positive control tissue: Cervix (Glandular cells, HPA High)
Negative control tissue: Ovary (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ARSB in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a nonimmune immunoglobulin control matched to the primary antibody’s host species and class; and ARSB knockout material, if available, as a biological negative (standard IHC practice). Quench endogenous peroxidase for chromogenic detection in kidney sections (standard IHC practice).
⚠️Feasibility: An ARSB-specific fixation window, fixation effect, and antigen-retrieval dependency are unreported in the supplied evidence; the A01609 tissue-IHC caption reports 2.5 µg/mL in human lung but does not state the fixative (caption: fixative not stated). The supplied evidence does not establish whether frozen sections or IF/ICC are easier for ARSB; HPA reports mainly Golgi localization in its ICC-IF images (HPA: approved Golgi location). Kidney tubules can contain endogenous biotin, which may mimic staining with avidin–biotin detection (standard IHC practice).
HPA tissue IHC evidence for ARSB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ARSB IHC Tips
Troubleshoot ARSB staining in paraffin sections by checking retrieval, compartment, controls, and scoring against the available IHC evidence.
How should I retrieve ARSB in paraffin sections with weak staining?
Start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval setting). Run a matched positive control alongside the study sections; human lung has a catalog antibody IHC image at 2.5 µg/mL, although its caption does not report fixation or retrieval (A01609 caption). If staining remains weak, compare a shorter citrate retrieval with an alternative buffer on adjacent sections, keeping antibody concentration and detection constant (general IHC optimization). Judge improvement by granular cytoplasmic signal and preserved tissue structure, rather than overall darkness alone (HPA tissue IHC profile; general IHC practice). Record the chosen retrieval conditions for every staining batch (general IHC practice).
Could fixation explain inconsistent ARSB staining across paraffin blocks?
ARSB-specific sensitivity to fixation is unknown from the supplied evidence; the catalog antibody’s human lung IHC caption gives 2.5 µg/mL but does not state a fixative (A01609 caption). Compare sections with documented processing histories in the same staining run, using the page’s citrate pH 6.0, 95–98 °C, 20 min retrieval consistently (page retrieval setting; general IHC practice). Keep section thickness, primary antibody concentration, and chromogen development consistent when assessing a processing effect (general IHC practice). Include a positive control on each run so a faint study section can be distinguished from a failed staining run (general IHC practice). Do not infer fixation sensitivity from ARSB glycosylation or tissue staining patterns (UniProt P15848; HPA tissue IHC profile).
Where should convincing ARSB staining appear in tissue sections?
Expect predominantly granular cytoplasmic staining in chromogenic sections, reflecting the reported tissue IHC profile (HPA tissue IHC profile). ARSB is annotated in lysosomes and at the cell surface, with no transmembrane segment; these annotations support evaluating intracellular granules and possible surface signal, but do not make every membrane outline specific (UniProt P15848 localisation and topology). ICC/IF images place ARSB mainly in the Golgi apparatus, a subcellular observation that may help interpret a concentrated perinuclear component without replacing tissue controls (HPA subcellular profile). Compare the pattern with high-staining glandular cells in cervix or kidney tubule cells on control sections (HPA tissue IHC). Treat isolated nuclear staining as suspect until supported independently (UniProt P15848 localisation; general IHC practice).
How can an unknown antibody epitope affect ARSB IHC interpretation?
ARSB has 2 annotated isoforms, while the supplied catalog caption does not define the antibody epitope or isoform coverage (UniProt P15848 isoforms; A01609 caption). Its precursor has a 1–36 signal peptide and a 37–533 annotated chain, so epitope placement matters when judging whether processing could change recognition (UniProt P15848 processing). Six annotated glycosylation sites and a modified residue at position 91 are additional reasons to verify epitope information before making a molecular explanation for staining differences (UniProt P15848 modifications). When epitope details are unavailable, assess staining with a second independently characterized antibody or a specific loss-of-signal control (general IHC validation practice). Do not assign a staining pattern to either isoform from these data alone (UniProt P15848 isoforms).
How should I plan an ARSB IF follow-up to the chromogenic IHC result?
Use the IF/ICC guide for the fluorescence experiment; the catalog evidence here is a tissue IHC image at 2.5 µg/mL, with no fixation stated, so it does not establish IF conditions (A01609 caption). Multiplex ARSB with a marker for the cell population under study and check each channel separately before interpreting overlap (general IF practice). Choose a far-red fluorophore where tissue autofluorescence compromises shorter wavelengths, and include unstained and single-color controls (general IF practice). Because ARSB is annotated in lysosomes and at the cell surface without a transmembrane segment, select permeabilisation according to the antibody epitope’s accessibility and verify that accessibility experimentally (UniProt P15848 localisation and topology; general IF practice). Compare any perinuclear concentration with the reported Golgi IF localisation (HPA subcellular profile).
What should I check when ARSB chromogenic staining looks diffuse or widespread?
ARSB can show widespread granular cytoplasmic staining, so broad tissue distribution alone is not evidence of background (HPA tissue IHC profile). Run a no-primary control and review endogenous peroxidase blocking before interpreting diffuse DAB deposition; these are general chromogenic IHC controls, not ARSB-specific findings (general IHC practice). Compare adjacent sections at the same 2.5 µg/mL catalog-image concentration only as an initial reference, since that caption provides no fixation or retrieval conditions (A01609 caption). Adjust blocking, antibody concentration, and chromogen development one variable at a time while retaining the page’s citrate pH 6.0 retrieval as the starting condition (page retrieval setting; general IHC optimization). Check whether background also appears in unstained edges or damaged tissue (general IHC practice).
How should I quantify heterogeneous ARSB staining in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis region before scoring, then record the percentage of positive cells and intensity separately (general IHC scoring practice). An H-score combines percentages at intensity grades 0–3 into a 0–300 scale; keep the same thresholds and chromogen development across sections (general IHC scoring practice). For spatial analyses, report positive cells per mm² and normalize to viable, evaluable tissue area, excluding folds, necrosis, and blank space (general IHC image analysis practice). Compare like cell populations because reported ARSB levels vary among tissue compartments, including high kidney tubule staining and undetected ovarian stromal staining (HPA tissue IHC). Record section batch and control performance beside each score (general IHC practice).
How do I distinguish true ARSB signal from chromogenic artefact?
Prioritize reproducible granular cytoplasmic signal in appropriate cells, consistent with the reported tissue IHC pattern; kidney tubule cells and cervical glandular cells are high-staining examples (HPA tissue IHC). ARSB is annotated in lysosomes and at the cell surface, whereas predominantly nuclear staining needs independent support (UniProt P15848 localisation; general IHC interpretation). Compare suspicious staining with a no-primary control and inspect section edges, necrotic areas, and regions prone to endogenous enzyme signal before calling cells positive (general chromogenic IHC practice). Interpret weak or absent staining cautiously because the tissue IHC evidence is approved with medium staining-to-RNA consistency and pending external verification (HPA tissue IHC reliability). Confirm consequential patterns with an independent specificity control (general IHC validation practice).
Boster reagents
Best ARSB / Arylsulfatase B IHC Antibodies
A01609 has ARSB IHC and IF images from human lung tissue (catalog: image captions). Human, Mouse and Rat are listed as reactive species (catalog: reactivity).
Real IHC data
Anti-Arylsulfatase B ARSB Antibody
Cat # A01609
A01609 is listed for IHC-P and IF, with a human lung IHC image at 2.5 μg/mL (catalog: applications; IHC image caption). Its IF image uses human lung tissue at 20 μg/mL; IHC and IF validation is reported only for human samples (catalog: IF image caption; dilution_raw).
Which to pick: For paraffin-section tissue IHC, pick A01609: IHC-P is listed, and its own image shows human lung staining at 2.5 μg/mL; the fixative is unreported (catalog: applications; IHC image caption). For IF, pick A01609 based on its human lung image at 20 μg/mL; ICC validation is unreported (catalog: IF image caption; applications). For Mouse or Rat, A01609 lists reactivity, but tissue IHC and IF validation is reported only for human samples; clonality is unreported (catalog: reactivity; dilution_raw; clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P15848 (ARSB_HUMAN, Arylsulfatase B).
- Human Protein Atlas. ARSB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ARSB subcellular location (ICC-IF): Mainly localized to the Golgi apparatus..
- Human Protein Atlas. ARSB antibody validation summary (2 antibodies).
- Differential effects of estrogen exposure on arylsulfatase B, galactose-6-sulfatase, and steroid sulfatase in rat prostate development. The Journal of steroid biochemistry and molecular biology 2014 — PMC4125552.
- Interaction of arylsulfatases A and B with maspin: A possible explanation for dysregulation of tumor cell metabolism and invasive potential of colorectal cancer. World journal of clinical cases 2019 — PMC6906558.
- Arylsulfatase B (N-acetylgalactosamine-4-sulfatase): potential role as a biomarker in prostate cancer. Prostate cancer and prostatic diseases 2013 — PMC3763935.
- Impact of salt exposure on N-acetylgalactosamine-4-sulfatase (arylsulfatase B) activity, glycosaminoglycans, kininogen, and bradykinin. Glycoconjugate journal 2013 — PMC3866634.
- PubMed PMID:2303452 — UniProt-cited evidence.
- PubMed PMID:1968043 — UniProt-cited evidence.
- PubMed PMID:7687847 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ARSB (Arylsulfatase B). Expected localisation Granular cytoplasm (HPA tissue IHC); lysosome and cell surface expected (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A01609 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Glandular cells); negative Ovary. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A01609); 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-ARSB, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Ovarian stroma may lack staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; signal peptide aa 1–36 is removed (UniProt); Regulation: Expression regulation is unreported (UniProt). Evidence: UniProt P15848, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus., Datasheet A01609, PMC4125552, PMC3866634. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ARSB/immunohistochemistry-ihc-guide.