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- Table of Contents
IDS / Iduronate 2-sulfatase · IHC design guide
Design Immunohistochemistry for IDS
Plan IDS staining in paraffin sections using the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet). Expect granular cytoplasmic staining, and compare high-staining colon glandular cells with adipocytes reported as undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IDS 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 (UniProt) | |
| Staining pattern | Granular cytoplasm in most tissues, including glandular cells (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 | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | No regulator specified in the supplied record (UniProt) | |
| Isoform / epitope | 3 isoforms; processed chains make epitope location relevant (UniProt) |
Section 1
Recommended IDS IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet); one published IDS IHC method uses free-floating mouse brain sections (PMC6517578).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA1917); 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-IDS, 0.5-1μg/ml (datasheet PA1917) |
| 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 | IDS-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 noteFor paraffin sections, start with citrate pH 6.0 at 95–98 °C for 20 min (page retrieval rule). The published free-floating method uses different conditions (PMC6517578).
Section 2
What Is the Expected IDS Staining Pattern?
IDS localises to lysosomes and has no transmembrane segment (UniProt P22304). HPA reports granular cytoplasmic IHC in most tissues, including high staining in colon glandular cells and cervical squamous cells (HPA tissue IHC). HPA rates the tissue IHC Approved but notes low consistency with RNA data and pending external verification (HPA tissue IHC).
| Granular cytoplasmic staining in colon glandular cells (HPA tissue IHC). | Expected IDS pattern; colon glandular staining is High (HPA tissue IHC). |
| Predominantly nuclear staining. | Unexpected for lysosomal IDS (UniProt P22304); assess background or off-target staining. |
| Strong staining in adipocytes. | Unexpected: adipocyte staining was not detected (HPA tissue IHC); check specificity. |
| Diffuse staining across cells and surrounding tissue. | Possible background; IDS is lysosomal (UniProt P22304). |
| No signal in colon glandular cells. | Unexpected relative to High staining (HPA tissue IHC); check assay performance. |
💡Expected IDS appearanceA positive IHC result shows high granular cytoplasmic staining in colon glandular or cervical squamous cells (HPA tissue IHC); predominantly nuclear staining is suspect given lysosomal localisation (UniProt P22304).
| Antibody validation (HPA tissue IHC) | IHC is Approved, with low RNA–staining consistency; external verification is pending (HPA tissue IHC). |
| Proteolytic processing (UniProt P22304) | Mature IDS has associated chains; check the antibody's epitope when interpreting staining. |
| Isoforms (UniProt P22304) | IDS has 3 isoforms; antibody recognition across them is unspecified. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in colon glandular cells (High; HPA tissue IHC). | Retrieval or detection may be inadequate (standard IHC practice). | Check controls; optimise retrieval and antibody dilution (standard IHC practice). |
| Predominantly nuclear signal. | Possible off-target staining; IDS is lysosomal (UniProt P22304). | Compare with a no-primary control and assess the staining pattern (standard IHC practice). |
| Adipocytes stain strongly. | Possible nonspecific signal; adipocytes were not detected (HPA tissue IHC). | Compare with a no-primary control and review antibody specificity (standard IHC practice). |
| Widespread diffuse chromogen. | Possible nonspecific binding or endogenous detection activity (standard IHC practice). | Check blocking and no-primary controls (standard IHC practice). |
| Expected cell types stain, but signal is hard to score. | Background may obscure granular cytoplasmic signal (HPA tissue IHC). | Adjust blocking and detection conditions using controls (standard IHC practice). |
| IF/ICC: what pattern should be expected? | HPA lists no ICC-IF images or subcellular location (HPA subcellular). | Use lysosomal localisation as the expectation (UniProt P22304); validate the IF signal independently. |
Experiment evidence
IHC & IF images showing IDS staining
PA1917 · IHCAnti-Iduronate 2 sulfatase antibody, PA1917, IHC(P) IHC(P): Human Placenta Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000010404-IDS/tissue/cerebral+cortex#imid_20628723
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000010404-IDS/tissue/colon#imid_20628641
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000010404-IDS/tissue/liver#imid_20628658
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000010404-IDS/tissue/kidney#imid_20628643
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IDS IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, non-glandular cells without specific signal should resemble the no-primary background, but do not assume every non-glandular cell is IDS-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for IDS; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus IDS-knockout material or a validated peptide-block control (standard IHC practice). For chromogenic colon IHC, quench endogenous peroxidase and check for background from inflammatory cells (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the PA1917 paraffin-section placenta caption does not state a fixative (PA1917 caption). Retrieval dependency is unreported; optimize antigen retrieval for the catalog antibody and evaluate punctate intracellular staining consistent with lysosomal IDS (UniProt P22304: lysosome; standard IHC practice). Frozen sections or IF cannot be judged easier from the supplied evidence; for colon IHC, assess inflammatory-cell peroxidase background with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for IDS
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced IDS IHC Tips
Troubleshoot IDS staining in paraffin sections by checking retrieval, cell type, lysosomal pattern and controls before interpreting chromogenic signal.
How should I retrieve IDS antigen when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then compare the same tissue across a small antibody dilution series so retrieval and antibody concentration are not changed simultaneously (general IHC practice). IDS is a lysosomal glycoprotein with 7 annotated glycosylation sites, so epitope accessibility may depend on processing, although this record does not establish a preferred alternative retrieval condition (UniProt P22304). If staining remains weak, test an alternative buffer or pH on matched sections while retaining the citrate condition as the reference (general IHC practice).
Could fixation explain weak or uneven IDS staining?
Target-specific sensitivity of IDS staining to fixation is unknown from the supplied evidence; the PA1917 placenta IHC caption does not state a fixative (caption PA1917). Record the fixative, fixation duration and processing history for every paraffin block, then compare sections processed together before changing the antibody concentration (general IHC practice). For routine formalin-fixed material, compare retrieval performance on matched sections and include a known staining control on each run (general IHC practice). Do not infer a particular fixation effect from IDS glycosylation or lysosomal localisation, or from HPA tissue staining patterns; neither establishes fixation sensitivity (UniProt P22304; HPA tissue IHC).
Where should convincing IDS staining appear in paraffin sections?
Look for granular cytoplasmic staining consistent with lysosomes, rather than using diffuse colour alone as evidence of IDS (UniProt P22304 localisation; HPA tissue IHC). IDS has a signal peptide and no annotated transmembrane segment, so a crisp plasma-membrane outline would need independent validation before assignment to this target (UniProt P22304 topology). HPA reports high staining in several epithelial or glandular cell populations, including colon glandular cells, but its tissue IHC reliability is Approved with low RNA–staining consistency and pending external verification (HPA tissue IHC). Compare the expected cellular pattern with a negative reagent control and matched sections before scoring (general IHC practice).
Can one IHC antibody distinguish IDS isoforms or processed chains?
Do not assign an IHC signal to a particular IDS isoform without an epitope map and isoform-specific validation: the record lists 3 isoforms but supplies neither for this antibody (UniProt P22304; supplied antibody evidence). IDS is processed from a 550-residue precursor into annotated 42 kDa and 14 kDa chains that remain associated in the active mature form (UniProt P22304). An antibody epitope within one chain could therefore affect what processing states are detectable, but the supplied caption does not identify the PA1917 epitope (UniProt P22304; caption PA1917). Interpret chromogenic staining as antibody-detected IDS immunoreactivity until specificity for an isoform or processing state is demonstrated (general IHC practice).
How should I investigate IDS by IF alongside the IHC result?
Treat IF as a separate validation experiment: the supplied PA1917 evidence is tissue IHC in placenta, with no stated fixative, and HPA lists no ICC/IF images (caption PA1917; HPA subcellular). Multiplex IDS with a marker identifying the expected cell population, and assess whether its signal is granular and cytoplasmic within those cells (HPA tissue IHC; general IF practice). Choose spectrally separated fluorophores after checking tissue autofluorescence, and include single-stain and no-primary controls when evaluating overlap (general IF practice). Because IDS is lysosomal and has no transmembrane segment, select permeabilisation appropriate for an intracellular epitope, then verify access without assuming the antibody's epitope is known (UniProt P22304; general IF practice).
What should I check when IDS DAB staining is widespread?
First compare the stained section with a no-primary control and examine whether colour follows tissue edges, folds or damaged areas (general IHC practice). Block endogenous peroxidase before chromogenic detection and check the DAB development time, because either step can contribute to apparent signal unrelated to antibody binding (general IHC practice). Titrate the IHC-validated antibody and detection reagent on matched sections, retaining a section with interpretable granular cytoplasmic staining as a run control (general IHC practice; HPA tissue IHC). Widespread smooth membrane or nuclear colour is discordant with the supplied lysosomal localisation and should not be scored as confirmed IDS (UniProt P22304).
How can I quantify IDS IHC without conflating cell composition and intensity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then report the percentage of positive cells and an intensity-based H-score using fixed thresholds across matched sections (general IHC practice). If positive cells are sparse, also report their density per mm² of viable tissue and state how excluded areas were handled (general IHC practice). Normalise comparisons to the number of evaluable cells or viable tissue area within the same annotated compartment, and retain the same counterstain and imaging settings (general IHC practice). Score granular cytoplasmic signal separately from diffuse background; HPA describes that pattern but flags low consistency with RNA expression (HPA tissue IHC).
How do I distinguish genuine IDS signal from staining artefacts?
Give greatest weight to reproducible granular cytoplasmic staining in an appropriate cell population, consistent with IDS lysosomal localisation and the HPA tissue pattern (UniProt P22304; HPA tissue IHC). Check whether apparent positives are confined to section edges, folds or necrotic regions, and compare them with no-primary controls for nonspecific colour (general IHC practice). Review the peroxidase block when DAB colour persists without primary antibody, since endogenous peroxidase can generate chromogenic signal (general IHC practice). Treat unexpected nuclear or sharply membrane-restricted staining cautiously, and do not call a negative cell type biologically IDS-free from IHC alone; HPA's tissue staining has low RNA–staining consistency and awaits external verification (UniProt P22304; HPA tissue IHC).
Boster reagents
Best IDS / Iduronate 2-sulfatase IHC Antibodies
PA1917 has human tissue IHC data from a paraffin section of placenta (catalog image caption); no IF data are supplied (catalog applications and images).
Real IHC data
Anti-Iduronate 2 sulfatase/IDS Antibody ®
Cat # PA1917
PA1917 is listed for human IHC (catalog applications and reactivity). Its IHC image shows a human placenta paraffin section (catalog image caption); no IF image is supplied (catalog images).
Which to pick: Choose PA1917 for human tissue IHC in paraffin sections (catalog reactivity and image caption); the fixative is unreported (catalog image caption). No listed antibody is validated for IF/ICC or other species (catalog applications and reactivity). PA1917 is a rabbit antibody with no clone specified (catalog host and clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P22304 (IDS_HUMAN, Iduronate 2-sulfatase).
- Human Protein Atlas. IDS tissue IHC expression (reliability: Approved).
- Human Protein Atlas. IDS subcellular location (ICC-IF): Highest expression in HTERT-RPE1: 92.0 nTPM.
- Human Protein Atlas. IDS antibody validation summary (1 antibodies).
- Identification of the iduronate-2-sulfatase proteome in wild-type mouse brain. Heliyon 2019 — PMC6517578.
- Targeting Brain Disease in MPSII: Preclinical Evaluation of IDS-Loaded PLGA Nanoparticles. International journal of molecular sciences 2019 — PMC6514713.
- The relevance of prelamin A and RAD51 as molecular biomarkers in cervical cancer. Oncotarget 2017 — PMC5706871.
- Human cytomegalovirus in high grade serous ovarian cancer possible implications for patients survival. Medicine 2018 — PMC5794372.
- PubMed PMID:2122463 — UniProt-cited evidence.
- PubMed PMID:8244397 — UniProt-cited evidence.
- PubMed PMID:8717057 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IDS (Iduronate 2-sulfatase). Expected localisation Granular cytoplasm (HPA tissue IHC); lysosome (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PA1917 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA1917); 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-IDS, 0.5-1μg/ml (datasheet PA1917); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; processed chains make epitope location relevant (UniProt); Regulation: No regulator specified in the supplied record (UniProt). Evidence: UniProt P22304, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in HTERT-RPE1: 92.0 nTPM, Datasheet PA1917, PMC6517578. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IDS/immunohistochemistry-ihc-guide.