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SUMO3 / Small ubiquitin-related modifier 3 · IHC design guide
Design Immunohistochemistry for SUMO3
Plan SUMO3 staining in paraffin sections around the predominantly nuclear tissue pattern (HPA tissue IHC). The guide covers fixation, staining controls and interpretation, including possible SUMO2/3 cross-reactivity (HPA tissue IHC; datasheet: IHC image caption).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SUMO3 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 | Predominantly nuclear in tissue IHC (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adipose tissue+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 formalin fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | SUMO2/3 cross-reactivity may confound staining (HPA tissue IHC; datasheet: IHC image caption) | |
| Regulation | Conjugation requires SAE1–SAE2 and UBE2I (UniProt) | |
| Isoform / epitope | 2 isoforms; processing removes residues 93–103; check epitope (UniProt) |
Section 1
Recommended SUMO3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 2 published paraffin-section IHC examples (PMC11345958; PMC11496881).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A05193); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-SUMO3, 5 μg/mL (datasheet A05193) |
| 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 | SUMO3-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule: nuclear antigen; UniProt P55854: nucleus).
Section 2
What Is the Expected SUMO3 Staining Pattern?
SUMO3 is reported in the nucleus, cytoplasm and PML bodies, and has no transmembrane segment (UniProt P55854). In tissue IHC, expect mainly nuclear staining across many cell types, including adipocytes and respiratory epithelial cells (HPA: nuclear expression in most tissues; High in adipocytes and respiratory epithelial cells). HPA rates the tissue pattern Supported, with medium agreement between staining and RNA data, and cautions that the antibodies may detect protein from more than one gene (HPA: reliability description).
| Distinct nuclear staining in adipocytes or bronchial respiratory epithelial cells. | Consistent with the reported IHC pattern and High staining in those cells (HPA: nuclear expression; High in adipocytes and respiratory epithelial cells). Compare nuclei with the surrounding tissue before scoring intensity; HPA's cross-gene caution limits SUMO3-specific attribution (HPA: reliability description). |
| Strong staining confined to cell membranes, with little nuclear staining. | Treat this as a compartment mismatch requiring investigation: SUMO3 has no transmembrane segment (UniProt P55854 topology), while HPA reports predominantly nuclear tissue staining (HPA: tissue IHC profile). Cytoplasmic signal alone is less decisive because UniProt also lists cytoplasmic localisation (UniProt P55854 subcellular location). |
| The strongest signal is in a cell population outside the expected pattern for the selected reference tissue. | Check cell identity and compare with the relevant HPA image; the supplied HPA list is not an exhaustive map of negative cell types (HPA: tissue IHC profile; no negative entries). Unexpected staining can suggest cross-reactivity or endogenous detection activity, but its appearance alone does not establish either cause. |
| Brown colour spreads across nuclei, cytoplasm and extracellular areas without clear cell boundaries. | Treat diffuse signal as background until controls show a defined pattern. HPA reports nuclear expression in most tissues (HPA: tissue IHC profile), but this does not validate indiscriminate colour. Review blocking, washing, chromogen development and the no-primary control as general IHC checks. |
| No convincing signal in a reference area expected to stain strongly. | A blank adipocyte or bronchial epithelial reference area conflicts with HPA's High staining entries (HPA: High in adipocytes and respiratory epithelial cells). First confirm that the expected cells are present and that the detection control worked; a blank section alone cannot distinguish a technical failure from antibody-specific behaviour. |
💡Expected SUMO3 appearanceCall a positive result when identifiable cells show clear, mainly nuclear chromogen, with strong reference staining where HPA reports High levels (HPA: nuclear expression; High in adipocytes and respiratory epithelial cells); widespread extracellular colour or membrane-only staining is suspect against that pattern (HPA: tissue IHC profile; UniProt P55854 topology).
| Compartment used for scoring | Prioritise nuclear localisation in paraffin tissue because that is the observed IHC profile (HPA: nuclear expression in most tissues). UniProt also lists cytoplasm and PML bodies (UniProt P55854 subcellular location); the supplied tissue profile does not assign a separate expected IHC intensity to either. |
| Choice of positive reference | Adipocytes, hematopoietic cells and bronchial respiratory epithelial cells are reported High, while cardiomyocytes and smooth muscle cells are reported Low (HPA: tissue IHC positive and low lists). Compare the same identified cell type within a section; Low is not a validated negative control. |
| Antibody specificity | HPA describes the tissue evidence as Supported with medium staining–RNA agreement and warns that the antibodies target protein from more than one gene (HPA: reliability description). A matching nuclear pattern therefore supports interpretation of the stain, but cannot by itself establish that every stained molecule is SUMO3. |
| SUMO3 processing and variants | The record lists two isoforms and processing from a 103-residue precursor to a 1–92 chain (UniProt P55854 isoforms and processing). Epitope placement and variant coverage are not supplied, so do not infer that a chosen antibody detects both isoforms or the removed segment. |
| Conjugated SUMO3 | SUMO3 can attach covalently to target proteins as a monomer or polymer (UniProt P55854 function). That biology is compatible with staining at multiple intracellular sites, but tissue IHC localisation alone cannot identify a conjugated substrate or separate conjugated from unconjugated SUMO3. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclear staining is weak in a High reference cell type. | The reference expectation may be unmet, or detection may be weak; HPA reports High staining in adipocytes and bronchial respiratory epithelial cells (HPA: tissue IHC positive list). | Confirm the cells and section quality, then review the catalog antibody's IHC-P instructions, retrieval settings and detection control. Treat retrieval adjustments as general IHC optimisation; no SUMO3-specific fixation sensitivity is established here. |
| Colour persists in the no-primary control. | Endogenous detection activity or nonspecific reagent deposition can produce chromogen without primary antibody; this is a general chromogenic IHC failure mode. | Check the detection system's endogenous activity block, blocking and wash steps. Compare the no-primary and primary-stained sections before assigning the colour to SUMO3. |
| Staining is diffuse across the section. | Excess background can obscure the expected mainly nuclear pattern (HPA: tissue IHC profile); the appearance does not identify a single cause. | Inspect background controls and tissue edges, then optimise the catalog antibody's IHC-P dilution, washing and chromogen development within its instructions. Score only cells whose boundaries and nuclei remain interpretable. |
| Membrane-only staining dominates. | That compartment conflicts with the reported mainly nuclear tissue pattern (HPA: tissue IHC profile) and lack of a transmembrane segment (UniProt P55854 topology). | Check the no-primary control, cell boundaries and antibody identification. Repeat against an HPA High reference cell type before treating membrane colour as target staining (HPA: tissue IHC positive list). |
| A Low reference cell type shows visible nuclear colour. | Low does not mean absent: HPA lists cardiomyocytes and smooth muscle cells as Low, without defining them as negative controls (HPA: tissue IHC low list). | Compare intensity and compartment with a High reference on a comparable run (HPA: tissue IHC positive and low lists). Do not call low-level nuclear colour a false positive solely because the reference is labelled Low. |
| What should an IF/ICC image show? | HPA reports supported nucleoplasm and nuclear-body localisation in ICC-IF, with additional approved localisation in sperm structures (HPA: subcellular summary and locations). | Look for nucleoplasmic and nuclear-body signal when interpreting IF/ICC (HPA: supported subcellular locations). Use the separate IF/ICC guide for that application; the supplied IHC evidence does not establish an IF/ICC protocol. |
Experiment evidence
IHC & IF images showing SUMO3 staining
A05193 · IHCImmunohistochemistry of SUMO2/3 in rat liver tissue with SUMO2/3 antibody at 5 μg/mL.
A05193 · IFImmunofluorescence of SUMO3 in mouse liver tissue with SUMO3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184900-SUMO3/tissue/cerebral+cortex#imid_12571141
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184900-SUMO3/tissue/colon#imid_14433782
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184900-SUMO3/tissue/liver#imid_12571076
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184900-SUMO3/tissue/kidney#imid_12571077
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB012180; green = SUMO3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB012180; green = SUMO3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SUMO3 IHC & IF
🧪Run adipose tissue first and confirm staining in adipocytes (HPA: High in adipocytes). HPA detects SUMO3 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat any unstained cells within the positive section as background references rather than proven SUMO3-negative cells (HPA: detected in all 45 scored tissues).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: SUMO3 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 SUMO3 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Nucleoplasm (supported), Nuclear bodies (supported) (HPA subcellular).
Technical controls: Run a secondary-only control and an irrelevant immunoglobulin control matched to the primary antibody’s host species, isotype and clonality; use SUMO3 knockout material, if available, as a biological specificity control (standard IHC practice). Quench endogenous peroxidase before chromogenic detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the A05193 rat-liver SUMO2/3 IHC caption leaves the fixative unreported (caption: fixative not stated). Antigen-retrieval dependence is unreported, and the evidence does not establish whether frozen sections or IF would be easier; ICC-IF images support nucleoplasm and nuclear-body localization as a reference for IF interpretation (HPA: supported main locations). In adipose sections, extracted lipid can leave large empty adipocyte spaces, so assess staining in the remaining cellular compartment (standard paraffin histology; HPA: High in adipocytes).
HPA tissue IHC evidence for SUMO3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SUMO3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SUMO3 IHC Tips
Use nuclear localisation and controlled chromogenic staining to assess SUMO3, while accounting for antibody cross-reactivity with SUMO2 (UniProt P55854; HPA tissue IHC).
How should I optimise retrieval when nuclear SUMO3 staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Cool sections in retrieval buffer, then compare test and control sections processed in the same run so heating differences do not masquerade as expression differences (standard IHC practice). If staining remains weak, test a shorter or longer retrieval time on adjacent sections while keeping buffer and pH constant (standard IHC practice). Score nuclear signal and tissue preservation together; strong diffuse colour after harsher retrieval does not by itself establish specific SUMO3 detection (HPA subcellular; standard IHC practice).
Could fixation explain weak staining in my paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the rat liver caption reports staining at 5 µg/mL but does not state a fixative (A05193 tissue-IHC caption). Record fixative, fixation duration, section thickness and storage for each specimen before attributing a weak result to fixation (standard IHC practice). For newly collected material, process matched samples consistently and compare adjacent sections after the same pH 9.0, 20 min retrieval step (page retrieval rule; standard IHC practice). Assess nuclear staining alongside morphology and a known positive control, since either processing variation or low detectable antigen can reduce signal (HPA tissue IHC; standard IHC practice).
Should chromogenic staining appear only in nuclei?
Prioritise nuclear staining when reading tissue sections: the tissue profile describes nuclear expression in most tissues, and cell imaging supports nucleoplasm and nuclear bodies (HPA tissue IHC; HPA subcellular). UniProt also lists cytoplasmic localisation and PML bodies, so a cytoplasmic component is possible but needs its own specificity checks (UniProt P55854 subcellular). SUMO3 has no transmembrane segment, making a crisp plasma-membrane-only pattern difficult to reconcile with its recorded localisation (UniProt P55854 topology and subcellular). Compare the nuclear and cytoplasmic compartments separately at matched exposure to chromogen, and verify an unexpected pattern with controls and an independently characterised antibody (standard IHC practice).
Can this antibody distinguish SUMO3 isoforms or SUMO2 in tissue?
Do not assign a chromogenic signal to one SUMO3 isoform without epitope and validation data: the record lists 2 isoforms, but supplies no antibody epitope (UniProt P55854 isoforms; supplied product evidence). The selected caption calls the reagent a SUMO2/3 antibody, and the tissue profile cautions that staining targets protein from more than one gene (A05193 tissue-IHC caption; HPA tissue IHC). SUMO3 is processed from a 103-residue precursor to a 1–92 mature chain, so epitope position can affect which forms are detected (UniProt P55854 processing). Use an epitope map and an orthogonal specificity control before reporting isoform-specific or SUMO3-only staining (standard IHC practice).
How can IF help check the cell source and compartment of IHC signal?
On a matched section, multiplex SUMO2/3 with a marker of the expected cell type, such as a hepatocyte marker when examining the captioned rat liver, and compare staining within marked cells (A05193 tissue-IHC caption; standard IF practice). Choose a far-red fluorophore where tissue autofluorescence obscures shorter wavelengths, and inspect an unstained or secondary-only section in each channel (standard IF practice). Because the principal expected signal is nucleoplasmic or in nuclear bodies, use a mild permeabilisation step that admits antibody to the nucleus, such as 0.1% Triton X-100 for 5 min, then check morphology (HPA subcellular; standard IF practice). Treat agreement with IHC as supporting localisation, while retaining the SUMO2/3 specificity caveat (A05193 tissue-IHC caption; HPA tissue IHC).
What should I check if DAB stains much of the section?
First inspect a no-primary control for endogenous peroxidase activity and detection-reagent background, then check whether DAB development was excessive (standard chromogenic IHC practice). Apply a peroxidase block before detection and optimise protein blocking and primary-antibody concentration using matched sections; the caption reports 5 µg/mL for rat liver but gives no paraffin processing details (standard IHC practice; A05193 tissue-IHC caption). Compare background in tissue edges, damaged regions and intact central tissue rather than treating all brown deposits as cellular signal (standard IHC practice). Specificity is more convincing when staining resolves to the expected nuclear compartments and the no-primary control stays clean (HPA tissue IHC; HPA subcellular; standard IHC practice).
How should I score SUMO3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear compartment before scoring, since the tissue profile reports predominantly nuclear expression (HPA tissue IHC). For each case, record the percentage of positive nuclei and an H-score using intensity grades 0–3; use positive nuclei per mm² when cell density itself is the outcome (standard IHC quantification practice). Normalise positive counts to all evaluable nuclei in the same annotated compartment, or to measured tissue area for density, while excluding folds and necrosis (standard IHC quantification practice). Keep retrieval, antibody concentration, DAB development and scoring thresholds consistent across batches, and report the SUMO2/3 cross-reactivity limitation with the results (page retrieval rule; A05193 tissue-IHC caption; HPA tissue IHC).
How do I separate genuine staining from artefact?
A credible result should show cellular nuclear staining consistent with the reported tissue pattern and supported nucleoplasm or nuclear-body localisation (HPA tissue IHC; HPA subcellular). Treat isolated membrane-only colour cautiously because SUMO3 has no transmembrane segment, and review unexpected cell populations against morphology and a cell-type marker (UniProt P55854 topology; standard IHC practice). Exclude staining confined to section edges, necrotic areas or the no-primary control, which can indicate processing artefact or endogenous detection activity (standard IHC practice). Finally, report the signal as SUMO2/3-reactive unless antibody-specific evidence separates the proteins; the selected caption and tissue profile both indicate that distinction is unresolved (A05193 tissue-IHC caption; HPA tissue IHC).
Boster reagents
Best SUMO3 / Small ubiquitin-related modifier 3 IHC Antibodies
A05193 has real IHC data from rat liver and IF data from mouse liver (IHC and IF captions); its listed reactivity covers human, mouse and rat (catalog reactivity).
Real IHC data
Anti-SUMO3 Antibody
Cat # A05193
A05193 is listed for IHC-P and IF (catalog applications). Its images show rat liver IHC at 5 μg/mL and mouse liver IF at 20 μg/mL (IHC and IF captions).
Which to pick: For paraffin-section IHC, choose A05193 (catalog: IHC-P; IHC caption: rat liver at 5 μg/mL); the fixative is unreported in the caption. For IF/ICC planning, A05193 has mouse liver IF evidence at 20 μg/mL (IF caption), while ICC validation and clonality are unreported (catalog applications and clone field). For work across species, A05193 lists human, mouse and rat reactivity (catalog reactivity); the pictured IHC and IF examples are rat and mouse, respectively (IHC and IF captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P55854 (SUMO3_HUMAN, Small ubiquitin-related modifier 3).
- Human Protein Atlas. SUMO3 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SUMO3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear bodies. In addition localized to the calyx, connecting piece and flagellar centriole..
- Human Protein Atlas. SUMO3 antibody validation summary (3 antibodies).
- SUMO3 inhibition by butyric acid suppresses cell viability and glycolysis and promotes gemcitabine antitumor activity in pancreatic cancer. Biology direct 2024 — PMC11345958.
- β-catenin-inhibited Sumoylation modification of LKB1 and fatty acid metabolism is critical in renal fibrosis. Cell death & disease 2024 — PMC11496881.
- Small ubiquitin-related modifier 2/3 interacts with p65 and stabilizes it in the cytoplasm in HBV-associated hepatocellular carcinoma. BMC cancer 2015 — PMC4603762.
- PubMed PMID:9119407 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:10830953 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SUMO3 (Small ubiquitin-related modifier 3). Expected localisation Predominantly nuclear in tissue IHC (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A05193 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. 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 A05193); 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 Tris-EDTA buffer, pH 9.0 , 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-SUMO3, 5 μg/mL (datasheet A05193); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Caveat: SUMO2/3 cross-reactivity may confound staining (HPA tissue IHC; datasheet: IHC image caption); Isoform / epitope: 2 isoforms ; processing removes residues 93–103; check epitope (UniProt); Regulation: Conjugation requires SAE1–SAE2 and UBE2I (UniProt). Evidence: UniProt P55854, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm and nuclear bodies. In addition…, Datasheet A05193, PMC11345958, PMC11496881. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SUMO3/immunohistochemistry-ihc-guide.