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DIS3 / Exosome complex exonuclease RRP44 · IHC design guide
Design Immunohistochemistry for DIS3
Plan chromogenic DIS3 IHC in paraffin sections using the catalog antibody's 2–5 μg/mL range (datasheet A01736-2). Assess mainly nuclear staining (HPA tissue IHC) and consider high-staining bronchial respiratory epithelium as a positive control (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DIS3 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 | Mainly nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread, mainly nuclear staining across tissue cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01736-2) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Nucleolar localisation differs between studies (UniProt) | |
| Regulation | Widely expressed (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope effects are unestablished (UniProt) |
Section 1
Recommended DIS3 IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet A01736-2). One published DIS3 IHC protocol uses paraffin sections of rectal adenoma and carcinoma (PMC3533962).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01736-2) |
| Fixation | Image fixative and duration unreported (datasheet A01736-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01736-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01736-2) |
| Primary antibody | Rabbit anti-DIS3, 2-5 μg/ml (datasheet A01736-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01736-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01736-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DIS3-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous mainly nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A01736-2). For rectal specimens, the published alternative is citrate pH 6 at 95°C for 25 min (PMC3533962).
Section 2
What Is the Expected DIS3 Staining Pattern?
DIS3 staining should be mainly nuclear across many cell types, with stronger staining in selected epithelial, glandular, and trophoblastic cells (HPA: ubiquitous mainly nuclear tissue IHC; HPA: listed High cells). Cytosolic signal is also supported by ICC/IF (HPA: supported cytosol). DIS3 has no transmembrane segment, so a membrane rim is not the expected pattern (UniProt Q9Y2L1 topology). HPA rates its tissue IHC profile Approved, with medium consistency against RNA expression data (HPA: tissue IHC reliability).
| Predominantly nuclear staining in respiratory epithelial cells of bronchus or squamous epithelial cells of cervix, with clear cell boundaries. | This matches the reported mainly nuclear tissue pattern and two High cell populations (HPA: tissue IHC profile; HPA: High in bronchus respiratory epithelium and cervix squamous epithelium). Assess the stain in identified cells rather than treating every part of the section as equally informative (general IHC practice). |
| Staining forms a crisp plasma-membrane rim, extracellular deposit, or isolated surface outline while nuclei are unstained. | That distribution conflicts with the mainly nuclear tissue pattern and with DIS3 having no transmembrane segment (HPA: tissue IHC profile; UniProt Q9Y2L1 topology). Review morphology and detection controls before calling it DIS3; the compartment mismatch alone does not identify the source of the stain (general IHC practice). |
| Strong staining appears in a population expected to be low, while a listed High population on the same run is weak or unstained. | For example, HPA reports low staining in lung alveolar cells and High staining in bronchial respiratory epithelial cells (HPA: tissue IHC levels). Misidentified cells, cross-reactivity, or endogenous detection activity are possible explanations; neither tissue is a universal negative or positive control for every specimen (general IHC practice; HPA: no negative tissues listed). |
| Color spreads over stroma, empty spaces, and several compartments without discernible nuclear enrichment. | Diffuse background makes the expected mainly nuclear pattern difficult to score (HPA: tissue IHC profile). Uneven blocking, insufficient washing, or excess detection reagent can produce nonspecific chromogenic signal; inspect a reagent-control section and the slide morphology before interpreting intensity (general IHC practice). |
| A well-preserved bronchus section shows no signal in its respiratory epithelial cells. | Those cells are listed as High, so absent staining warrants a run-level check (HPA: High in bronchus respiratory epithelial cells). Confirm that tissue and nuclei are present, then review retrieval, primary-antibody application, and chromogenic detection using the assay's controls (general IHC practice). One negative section does not establish that DIS3 is absent (HPA: Approved, medium consistency). |
💡Expected DIS3 appearanceCall a convincing positive when identifiable cells show predominantly nuclear staining, especially listed High epithelial or glandular populations; a membrane-only rim or diffuse tissue-wide color is suspect (HPA: tissue IHC profile and High cell levels; UniProt Q9Y2L1 topology; general IHC practice).
| Tissue and cell selection | DIS3 is widely expressed, while HPA reports different staining levels by cell type (UniProt Q9Y2L1 tissue specificity; HPA: tissue IHC levels). Use a listed High population to judge whether the run can reveal nuclear signal; HPA supplies no negative tissue here (HPA: High cells; HPA: negative list empty). |
| Nucleolar interpretation | UniProt reports both nucleolar localization and a study finding DIS3 excluded from nucleoli (UniProt Q9Y2L1 subcellular notes). HPA supports nucleoplasm as the main ICC/IF location (HPA: subcellular). Do not require nucleolar enrichment to accept a mainly nuclear IHC result (HPA: tissue IHC profile). |
| Isoforms and antibody epitope | UniProt lists two DIS3 isoforms and several domains, but the supplied records do not locate an antibody epitope or show isoform-specific IHC staining (UniProt Q9Y2L1 isoforms and domains; HPA: supplied antibody records). An unexpected pattern cannot be assigned to one isoform from these data. |
| Antibody validation | HPA039281 is Approved for IHC, whereas no IHC status is supplied for HPA058762; both have Enhanced ICC validation (HPA: antibody validation records). The tissue profile itself is Approved with medium consistency, so compare compartment and cell identity alongside intensity (HPA: tissue IHC reliability and profile). |
| Detection chemistry | Endogenous enzyme activity and nonspecific reagent binding can add chromogenic color, independently of DIS3 localization (general IHC practice). Blocking and reagent controls help distinguish these signals from the mainly nuclear tissue pattern (general IHC practice; HPA: tissue IHC profile). No DIS3-specific fixation sensitivity is reported in the supplied sources (UniProt Q9Y2L1; HPA: supplied records). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a listed High cell population. | The section or assay may have failed; absence alone does not establish loss of DIS3 (HPA: High cell levels; general IHC practice). | Check tissue preservation and cell identity, then review retrieval, antibody application, and detection controls for the IHC run (general IHC practice). |
| Strong cytoplasmic color obscures nuclei. | Cytosol is a supported additional ICC/IF location, but tissue IHC is mainly nuclear; excessive background is also possible (HPA: subcellular; HPA: tissue IHC profile; general IHC practice). | Compare nuclear enrichment with morphology and reagent controls before scoring; do not count diffuse color alone as a specific positive (general IHC practice). |
| Only the cell surface or extracellular material stains. | This conflicts with the mainly nuclear profile and absence of a transmembrane segment (HPA: tissue IHC profile; UniProt Q9Y2L1 topology). | Inspect the section for deposits, review control staining, and avoid assigning the surface signal to DIS3 without independent support (general IHC practice). |
| Many cell types stain equally, including a listed low-staining population. | Broad expression is plausible, but HPA reports cell-level differences; excess background or loss of contrast may mask them (UniProt Q9Y2L1 tissue specificity; HPA: tissue IHC levels; general IHC practice). | Score identified cell populations separately and compare their nuclear signal with the run's controls (general IHC practice). |
| Is nucleolar staining required for a positive call? | UniProt's nucleolar reports differ, and HPA supports nucleoplasm as the main ICC/IF location (UniProt Q9Y2L1 subcellular notes; HPA: subcellular). | Use predominant nuclear staining as the IHC criterion; describe nucleolar staining separately if visible (HPA: tissue IHC profile; general IHC practice). |
| Can an ICC/IF image settle an uncertain chromogenic IHC result? | HPA supports nucleoplasmic and additional cytosolic ICC/IF localization, while its tissue IHC profile is mainly nuclear (HPA: subcellular; HPA: tissue IHC profile). | Use ICC/IF as compartment context, then assess the IHC section with its own morphology and controls; consult the separate IF/ICC guide for that application (general IHC practice). |
Experiment evidence
IHC & IF images showing DIS3 staining
A01736-2 · IHCIHC analysis of DIS3 using anti-DIS3 antibody (A01736-2). DIS3 was detected in a paraffin-embedded section of human breast cancer tissue. He
A01736-2 · IHCIHC analysis of DIS3 using anti-DIS3 antibody (A01736-2). DIS3 was detected in a paraffin-embedded section of human breast cancer tissue. He
A01736-2 · IFIF analysis of DIS3 using anti-DIS3 antibody (A01736-2) and anti-Beta Tubulin antibody (M01857-3). DIS3 was detected in immunocytochemical s
A01736 · IHCImmunohistochemistry of DIS3 in mouse testis tissue with DIS3 antibody at 2.5 μg/ml.
A01736 · IFImmunofluorescence of DIS3 in mouse testis tissue with DIS3 antibody at 20 μg/ml.
A01736-1 · IFIF analysis of DIS3 using anti-DIS3 antibody (A01736-1) and anti-Beta Tubulin antibody (M01857-3). DIS3 was detected in immunocytochemical s
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000083520-DIS3/tissue/cerebral+cortex#imid_11526523
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000083520-DIS3/tissue/colon#imid_11526449
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000083520-DIS3/tissue/liver#imid_11526458
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000083520-DIS3/tissue/kidney#imid_11526451
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA039281; green = DIS3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA039281; green = DIS3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DIS3 IHC & IF
🧪Run bronchus first; its respiratory epithelial cells should stain strongly for DIS3 (HPA: High in bronchus respiratory epithelial cells). HPA detects DIS3 in all 45 scored tissues (HPA: no negative tissue rows), so use no-primary and isotype controls for the negative comparison; no cell population on the bronchus slide is established as DIS3-negative, and weakly stained neighboring cells should be treated only as internal contrast (HPA: no negative tissue rows).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: None in HPA: DIS3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DIS3 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control for the rabbit primary; and, if available, a DIS3-knockout specimen processed alongside the section as a biological negative (A01736-2 IHC caption: rabbit primary; standard IHC practice). Block endogenous peroxidase before HRP/DAB detection and check inflammatory cells in bronchus for residual background (A01736-2 IHC caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A01736-2 paraffin-section caption does not state a fixative (A01736-2 IHC caption: fixative not stated). That example used heat retrieval in EDTA pH 8.0 before staining, but whether retrieval is required for DIS3 is unreported (A01736-2 IHC caption). HPA shows nucleoplasmic and additional cytosolic signal by ICC-IF, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC (HPA: subcellular localization; A01736-2 IHC caption).
HPA tissue IHC evidence for DIS3
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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: DIS3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced DIS3 IHC Tips
Troubleshoot DIS3 staining in paraffin sections by checking retrieval, nuclear localisation, and matched controls before interpreting differences in signal (datasheet A01736-2; HPA tissue IHC).
Which retrieval condition should I try first for DIS3 paraffin IHC?
Start with heat-mediated EDTA retrieval at pH 8.0 for DIS3 paraffin IHC (datasheet A01736-2). The selected tissue image used 2 μg/ml primary antibody overnight at 4°C, followed by DAB detection (caption A01736-2). Keep heating, cooling, antibody incubation, and chromogen development consistent across matched sections while adjusting one condition at a time (standard IHC practice). If staining remains weak, check deparaffinisation and reagent performance, then compare an alternative retrieval condition on adjacent sections as a fallback; assess nuclear signal alongside background and tissue integrity (standard IHC practice; HPA tissue IHC).
How should I investigate variable DIS3 staining between paraffin specimens?
The selected image identifies a paraffin section but does not state its fixative, so DIS3-specific fixation sensitivity is unknown (caption A01736-2). Record each specimen’s fixative, delay before fixation, and fixation duration, then process matched sections together when investigating variation (standard IHC practice). Begin comparison with the documented EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody incubation overnight at 4°C (caption A01736-2). Inspect morphology and batch controls before changing retrieval or antibody concentration, and do not infer a fixation effect from DIS3 localisation or modifications (standard IHC practice; UniProt Q9Y2L1).
Should DIS3 staining be nuclear, nucleolar, or cytoplasmic?
Expect mainly nuclear staining in tissue, with nucleoplasmic localisation supported by subcellular imaging and additional cytosolic signal reported there (HPA tissue IHC; HPA subcellular). DIS3 is predominantly nuclear, but reports differ on whether it occupies or is excluded from nucleoli (UniProt Q9Y2L1; PubMed:12429849; PubMed:20531386). Score nuclear and cytoplasmic staining separately, using a nuclear counterstain and comparable chromogen development across sections (standard IHC practice). If apparent nucleolar dots or diffuse cytoplasmic colour dominate, compare adjacent sections and negative controls before assigning them to DIS3; the available localisation evidence does not establish either pattern as diagnostic in this assay (HPA subcellular; standard IHC practice).
Can this antibody distinguish DIS3 isoforms or modified epitopes in tissue?
DIS3 has 2 reported isoforms, while the supplied antibody image does not identify its recognised epitope or demonstrate isoform selectivity (UniProt Q9Y2L1; caption A01736-2). Its annotated domains include PINc, CSD1, CSD2, and RNB, and recorded modifications include acetylation and phosphorylation (UniProt Q9Y2L1). Those annotations alone cannot show whether fixation, retrieval, or a modification changes antibody binding in paraffin sections (UniProt Q9Y2L1; standard IHC practice). If an isoform-specific interpretation matters, obtain epitope mapping and use an independently validated comparator; report this stain as DIS3 immunoreactivity until specificity is established for that distinction (standard IHC practice).
How can I follow up the tissue IHC pattern with multiplex IF?
Treat IF/ICC as a separate assay while using the mainly nucleoplasmic and additional cytosolic distribution as localisation expectations (HPA subcellular). Pair DIS3 with a marker for the cell type being examined, and select spectrally separated fluorophores after checking tissue autofluorescence and single-colour controls (standard IF practice). DIS3 has no transmembrane segment and is reported inside cells, so include a permeabilisation step that gives the antibody access to intracellular epitopes; optimise its strength for morphology (UniProt Q9Y2L1; standard IF practice). Compare nuclear, cytosolic, and marker channels separately, and do not transfer the paraffin image’s fixation assumptions or DAB readout to IF (caption A01736-2; standard IF practice).
What should I check when DIS3 DAB staining is diffuse?
First compare the diffuse colour with a no-primary control and a matched section processed in the same run (standard IHC practice). The documented workflow used 10% goat serum blocking and a peroxidase-linked secondary with DAB, so review blocking, secondary binding, and chromogen development as possible contributors (caption A01736-2; standard IHC practice). Include a peroxidase block and inspect for endogenous enzyme signal, especially if colour persists without primary antibody (standard IHC practice). Adjust primary concentration or incubation only after checking reagent controls, and judge the result against the expected mainly nuclear pattern rather than total brown area (HPA tissue IHC; standard IHC practice).
How should I quantify DIS3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell compartments before scoring because the tissue profile is mainly nuclear, while cytosolic localisation is also reported (HPA tissue IHC; HPA subcellular). For nuclear staining, record the percentage of positive cells and intensity by category, then calculate an H-score on the 0–300 scale if that suits the study (standard IHC practice). For spatial comparisons, report positive cells per mm² of viable tissue and keep segmentation and DAB thresholds fixed across slides (standard IHC practice). Normalise counts to the number of eligible nuclei or viable tissue area, and document cell type, batch, and excluded damaged regions (standard IHC practice).
When is an unexpected DIS3 staining pattern likely to be artefact?
A convincing result should show reproducible staining in intact cells, with a mainly nuclear pattern consistent with the tissue profile (HPA tissue IHC; standard IHC practice). High signal in several epithelial cell populations is reported, but low staining in some other populations means cell identity matters when comparing fields (HPA tissue IHC). Treat edge accentuation, necrotic regions, and colour present in no-primary controls as possible processing or detection artefacts; persistent DAB signal may reflect endogenous peroxidase (standard IHC practice). Resolve predominantly extracellular or unexplained diffuse staining with matched controls and independent specificity evidence before interpreting it as altered DIS3 biology (UniProt Q9Y2L1; standard IHC practice).
Boster reagents
Best DIS3 / Exosome complex exonuclease RRP44 IHC Antibodies
DIS3 IHC images cover paraffin-embedded human breast and colon cancer sections (A01736-2 image captions) and mouse testis (A01736 image caption); IF images include HeLa cells (A01736-1 image caption).
Real IHC data
Anti-DIS3 Antibody ®
Cat # A01736-2
Real IHC data
Anti-DIS3 Antibody
Cat # A01736
Real IF data
Anti-DIS3 Antibody ®
Cat # A01736-1
A01736-2 has IHC images from human breast and colon cancer sections and an IF image from HeLa cells (catalog image captions); A01736 has IHC and IF images from mouse testis (catalog image captions). A01736-1 has IF images from HeLa cells and a paraffin-embedded human breast cancer section (catalog image captions).
Which to pick: For tissue IHC, choose A01736-2 for human paraffin sections (A01736-2 IHC captions: breast and colon cancer; datasheet: 2–5 μg/ml) or A01736 for mouse testis (A01736 IHC caption: 2.5 μg/ml); neither IHC caption reports a fixative (catalog IHC captions). For IF/ICC, A01736-1 has a HeLa cell image and lists both applications (A01736-1 IF caption; catalog applications). For cross-species IHC planning, A01736-2 lists human, mouse and rat reactivity, though its own IHC images show human sections (catalog reactivity; A01736-2 IHC captions); clone identity is unreported for all three (catalog clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y2L1 (RRP44_HUMAN, Exosome complex exonuclease RRP44).
- Human Protein Atlas. DIS3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DIS3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. DIS3 antibody validation summary (2 antibodies).
- Genomic profiling of rectal adenoma and carcinoma by array-based comparative genomic hybridization. BMC medical genomics 2012 — PMC3533962.
- PubMed PMID:9562621 — UniProt-cited evidence.
- PubMed PMID:11935316 — UniProt-cited evidence.
- PubMed PMID:10231032 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DIS3 (Exosome complex exonuclease RRP44). Expected localisation Mainly nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01736-2). Validated IHC antibody: A01736-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01736-2); 2. Fixation: Image fixative and duration unreported (datasheet A01736-2); 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: EDTA pH 8.0 (datasheet A01736-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01736-2); 8. Primary antibody: Rabbit anti-DIS3, 2-5 μg/ml (datasheet A01736-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01736-2). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Nucleolar localisation differs between studies (UniProt); Isoform / epitope: 2 isoforms; epitope effects are unestablished (UniProt); Regulation: Widely expressed (UniProt). Evidence: UniProt Q9Y2L1, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A01736-2, PMC3533962. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DIS3/immunohistochemistry-ihc-guide.