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SUMO2 / Small ubiquitin-related modifier 2 · IHC design guide
Design Immunohistochemistry for SUMO2
Plan paraffin-section SUMO2 IHC around the general nuclear staining seen in tissues (HPA tissue IHC). The guide covers fixation, chromogenic detection and interpretation of an antibody that detects SUMO2 and SUMO3 (datasheet M01282-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SUMO2 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 | General nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across many cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01282-2) | |
| Positive control | Adrenal gland+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 consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | SUMO3 cross-detection may confound scoring (datasheet M01282-2) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; residues 94–95 removed; epitope impact unknown (UniProt) |
Section 1
Recommended SUMO2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: M01282-2) is accompanied by published brain and mouse sciatic nerve IHC conditions (PMC10571773; PMC6524499).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M01282-2) |
| Fixation | Image fixative and duration unreported (datasheet M01282-2); 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 M01282-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01282-2) |
| Primary antibody | Rabbit monoclonal (clone IBG-19) anti-SUMO2, 1:50 (datasheet M01282-2) |
| Primary incubation | Overnight at 4 °C (datasheet M01282-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M01282-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SUMO2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval in EDTA pH 8.0 (datasheet: M01282-2); citrate pH 6.0 is a published alternative for mouse sciatic nerve (PMC6524499).
Section 2
What Is the Expected SUMO2 Staining Pattern?
SUMO2 is nuclear, including PML bodies, and has no transmembrane segment (UniProt P61956). In paraffin sections, expect predominantly nuclear staining across many cell types, with high staining in several glandular and epithelial populations (HPA tissue IHC). HPA calls the tissue pattern Supported but reports medium consistency with RNA and cautions that the staining may detect proteins from more than one gene (HPA tissue IHC).
| Predominantly nuclear staining in adrenal glandular cells, bone marrow hematopoietic cells, or bronchial epithelium. | This fits HPA's general nuclear pattern and its High staining calls for those cells (HPA tissue IHC). Compare intensity within the same staining run; a High call does not require every nucleus to look identical (HPA tissue IHC; standard IHC practice). |
| Strong, widespread cytoplasmic or membranous staining with little nuclear signal. | This conflicts with the reported nuclear localization and lack of a transmembrane segment (UniProt P61956; HPA tissue IHC). Review background, detection controls, and antibody specificity before interpreting it as SUMO2 distribution (standard IHC practice; HPA tissue IHC caution). |
| Intense staining in a cell population where HPA reports Low staining, such as liver cholangiocytes. | HPA's Low call makes this a discrepancy, not proof that the cells are negative (HPA tissue IHC). Check tissue identification and controls; cross-reactivity or endogenous detection activity are possible explanations (HPA tissue IHC caution; standard IHC practice). |
| Diffuse color over tissue and empty areas, obscuring nuclear boundaries. | The slide cannot be scored reliably for the expected nuclear pattern (UniProt P61956; standard IHC practice). Background can arise from detection reagents or incomplete blocking; compare a no-primary control and inspect the counterstain (standard IHC practice). |
| No nuclear signal in a section containing HPA High cells, such as breast glandular cells. | The result does not reproduce the reported High nuclear pattern (HPA tissue IHC). Check tissue preservation, the catalog antibody's IHC-P instructions, detection reagents, and an appropriate positive control before treating the tissue as SUMO2-negative (standard IHC practice). |
💡Expected SUMO2 appearanceCall a result positive when nuclei show clear chromogenic staining in an HPA High population, such as esophageal squamous epithelial cells; dominant cytoplasmic or membranous color is discordant with the reported pattern (HPA tissue IHC; UniProt P61956).
| Tissue and cell selection | SUMO2 is broadly expressed (UniProt P61956), while HPA reports High staining in selected cells and Low staining in others; a Low call is not a negative control (HPA tissue IHC). |
| Antibody specificity | HPA rates tissue staining Supported and warns that antibodies may target proteins from more than one gene; nuclear staining alone cannot prove SUMO2-specific detection (HPA tissue IHC). |
| Protein forms | UniProt lists 2 isoforms, a 95-residue precursor, and a mature chain spanning residues 1–93 (UniProt P61956). Epitope coverage is unspecified, so these facts cannot predict staining differences (UniProt P61956). |
| Detection chemistry | For enzyme-based chromogenic IHC, endogenous activity can mimic specific color; use the detection system's appropriate blocking and no-primary controls (standard IHC practice). |
| IF/ICC Q: What pattern should appear? | A: Mainly nucleoplasm and nuclear bodies, with additional sperm structures reported; HPA cautions that the antibodies target proteins from multiple genes (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclei are weak in an HPA High cell population. | The run may have insufficient signal; HPA High is a reference pattern, not a guaranteed result for each section (HPA tissue IHC; standard IHC practice). | Verify positive-control staining and follow the catalog antibody's IHC-P retrieval and dilution instructions (standard IHC practice). |
| Cytoplasmic color overwhelms nuclear staining. | The distribution conflicts with reported SUMO2 localization; nonspecific staining or detection background is possible (UniProt P61956; standard IHC practice). | Compare the no-primary control, then review blocking, antibody dilution, and detection conditions (standard IHC practice). |
| Color appears in the no-primary control. | The signal does not require primary antibody and may reflect endogenous detection activity or reagent background (standard IHC practice). | Apply the detection system's appropriate endogenous-activity block and check its reagents (standard IHC practice). |
| A nominally Low population stains strongly. | HPA's Low designation conflicts with strong staining but does not establish absence of SUMO2; antibody cross-detection remains possible (HPA tissue IHC). | Confirm the cell type and assess staining with an independent, appropriately validated antibody if available (standard IHC practice; HPA tissue IHC caution). |
| Nuclear color is indistinct despite visible tissue staining. | Diffuse background or heavy counterstaining can obscure nuclear boundaries (standard IHC practice). | Compare the no-primary control and adjust background control or counterstain according to the staining system (standard IHC practice). |
| Positive-control and test sections are both blank. | A shared staining-run failure is more plausible than a tissue-specific negative result (standard IHC practice). | Check section handling, retrieval, antibody application, and chromogen performance against the catalog IHC-P instructions (standard IHC practice). |
Experiment evidence
IHC & IF images showing SUMO2 staining
M01282-2 · IHCIHC analysis of SUMO2/3 using anti-SUMO2/3 antibody (M01282-2). SUMO2/3 was detected in a paraffin-embedded section of human lung cancer tis
M01282-2 · IHCIHC analysis of SUMO2/3 using anti-SUMO2/3 antibody (M01282-2). SUMO2/3 was detected in a paraffin-embedded section of rat brain tissue. Hea
A01282-1 · IHCImmunohistochemistry of SUMO2/3 in rat liver tissue with SUMO2/3 antibody at 5 μg/mL.
A01282-1 · IFImmunofluorescence of SUMO2/3 in rat liver tissue with SUMO2/3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000188612-SUMO2/tissue/cerebral+cortex#imid_8972338
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000188612-SUMO2/tissue/colon#imid_8972272
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000188612-SUMO2/tissue/liver#imid_8972273
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000188612-SUMO2/tissue/kidney#imid_8972274
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042123; green = SUMO2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042123; green = SUMO2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SUMO2 IHC & IF
🧪Start with breast tissue and assess staining in glandular cells (HPA: High in breast glandular cells). HPA detects SUMO2 in all 45 scored tissues, so there is no validated negative tissue; no cells within the positive slide are established as target-negative, and any unstained cells should show preserved morphology without nuclear chromogen while the no-primary and isotype sections establish background (HPA: no negative rows; UniProt P61956: broadly expressed, nuclear).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: SUMO2 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 SUMO2 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Nucleoplasm (supported), Nuclear bodies (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only section and a concentration-matched rabbit IgG isotype section, matching clonality if known; use SUMO2 knockout material as a biological specificity control where available (caption: rabbit primary; standard IHC practice). Check breast sections for endogenous peroxidase activity before interpreting DAB signal (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected M01282-2 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 for lung cancer tissue, but does not establish that retrieval condition for breast tissue (caption: EDTA pH 8.0; HPA: High in breast glandular cells). Frozen sections and IF cannot be judged easier from the supplied evidence; interpret nuclear staining with awareness that the HPA subcellular summary cautions that its ICC-IF localisation is based on antibodies targeting proteins from multiple genes (HPA: subcellular summary).
HPA tissue IHC evidence for SUMO2
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial 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: SUMO2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SUMO2 IHC Tips
Troubleshoot SUMO2 staining in paraffin sections by checking retrieval, nuclear localisation, antibody specificity and scoring controls.
Which retrieval conditions should I start with for SUMO2 staining in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M01282-2). The catalog antibody produced chromogenic staining in a paraffin-embedded human lung cancer section after this retrieval, followed by 1:50 primary antibody overnight at 4°C (datasheet M01282-2). If nuclear staining is weak, vary heating time on adjacent sections while holding antibody dilution and detection conditions constant, then compare signal with tissue damage and background (standard IHC practice). Record the retrieval duration actually used, because the caption specifies the buffer and pH but gives no heating time (datasheet M01282-2).
Could fixation explain weak or uneven SUMO2 staining?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (datasheet M01282-2). Document fixation conditions for each block and compare sections processed with matched retrieval and detection settings before attributing a difference to SUMO2 biology (standard IHC practice). Inspect section edges and tissue preservation, since uneven processing can complicate assessment of nuclear chromogenic staining (standard IHC practice; HPA: general nuclear expression). Use the catalog method’s EDTA retrieval at pH 8.0 as the starting condition, and assess a control section alongside each fixation group (datasheet M01282-2; standard IHC practice).
Where should convincing SUMO2 staining appear, and how should cytoplasmic signal be assessed?
Expect predominantly nuclear staining, potentially including nuclear bodies, because SUMO2 is annotated in the nucleus and PML bodies and HPA reports general nuclear expression (UniProt P61956 subcellular; HPA tissue IHC profile). HPA also supports nucleoplasmic and nuclear-body localisation, with a caution that its antibodies target proteins from multiple genes (HPA subcellular). Compare nuclear signal with adjacent cytoplasm in well-preserved cells, using the same counterstain and detection settings across sections (standard IHC practice). Treat diffuse cytoplasmic DAB as unresolved until omission and specificity controls exclude background; SUMO2 has no transmembrane segment, but that fact alone does not establish antibody specificity (UniProt P61956 topology; standard IHC practice).
Can this antibody distinguish SUMO2 from SUMO3 or detect only conjugated SUMO2?
The selected product is described as an anti-SUMO2/3 antibody, so its staining should not be scored as SUMO2-specific without independent validation (datasheet M01282-2). SUMO2 has 2 annotated isoforms and is processed from a 95-residue precursor to a 1–93 chain (UniProt P61956 isoforms and processing). It can be covalently attached to substrates as a monomer or polymer, making a nuclear signal compatible with several molecular states rather than proof of conjugation (UniProt P61956 function). Check the antibody’s stated epitope and perform appropriate specificity controls before assigning staining to an isoform, SUMO2 alone, or a particular conjugated species (standard IHC practice).
How can IF help evaluate a SUMO2 IHC pattern?
Use IF as a separate localisation check and multiplex SUMO2/3 with a marker of the cell population being assessed, so nuclear signal can be assigned to the intended cells (standard IF practice; datasheet M01282-2 specificity). Choose a fluorophore whose emission is separated from the tissue’s autofluorescence, and inspect single-stain controls before interpreting overlap (standard IF practice). SUMO2 has no transmembrane segment and is annotated in the nucleus and PML bodies; permeabilise sufficiently to access the nuclear epitope while preserving morphology (UniProt P61956 topology and subcellular; standard IF practice). Compare the resulting nuclear pattern with chromogenic IHC without transferring the paraffin-section caption’s conditions to IF (datasheet M01282-2; standard IF practice).
What controls help distinguish SUMO2 signal from chromogenic background?
The selected IHC example used 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, a peroxidase-conjugated secondary, and DAB development (datasheet M01282-2). Include a no-primary control and a peroxidase block, and inspect whether DAB appears where specific primary-dependent nuclear staining should be absent (standard IHC practice; HPA: general nuclear expression). If background persists, compare adjacent sections with reduced primary concentration or shorter DAB development while keeping retrieval and imaging settings documented (standard IHC practice). Evaluate tissue folds, damaged edges and pigment separately from intact nuclei before calling any dark deposit positive (standard IHC practice).
How should SUMO2 nuclear staining be scored across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysed cell population and tissue area before scoring, then report the percentage of positive nuclei and an intensity-based H-score if staining permits reliable intensity categories (standard IHC practice; HPA: general nuclear expression). An H-score can combine percentages at intensity grades 0–3, yielding a possible range of 0–300 (standard IHC practice). Normalise counts to eligible intact nuclei or sampled tissue area, and keep retrieval, DAB development and image settings consistent across groups (standard IHC practice). Because this catalog antibody is described as anti-SUMO2/3, label the measurement accordingly unless independent controls establish SUMO2-specific detection (datasheet M01282-2).
When does an apparent SUMO2-positive pattern warrant caution?
Give greatest weight to staining in intact nuclei, consistent with SUMO2’s nuclear and PML-body annotation and HPA’s general nuclear IHC profile (UniProt P61956 subcellular; HPA tissue IHC profile). Recheck staining confined to tissue edges, necrotic regions or no-primary controls, where processing artefacts or endogenous enzyme activity can mimic DAB signal (standard IHC practice). A predominantly nonnuclear pattern needs additional validation before assignment to SUMO2, while nuclear staining alone cannot identify a particular substrate or SUMO2 conjugation state (UniProt P61956 subcellular and function; standard IHC practice). Interpret any positive result as SUMO2/3-reactive unless independent specificity evidence distinguishes the proteins (datasheet M01282-2).
Boster reagents
Best SUMO2 / Small ubiquitin-related modifier 2 IHC Antibodies
IHC images show paraffin-embedded human cancer and rat brain, plus rat liver (catalog IHC captions); an IF image shows rat liver (A01282-1 IF caption).
Real IHC data
Anti-Sumo2/3 Rabbit Monoclonal Antibody
Cat # M01282-2
Real IHC data
Anti-SUMO2/3 Antibody
Cat # A01282-1
M01282-2 lists IHC, IF and ICC for human, mouse and rat; its IHC images show paraffin-embedded human lung, liver and rectal cancers and rat brain (catalog applications/reactivity; M01282-2 IHC captions). A01282-1 lists IHC-P and IF for human, mouse and rat, with IHC and IF images from rat liver (catalog applications/reactivity; A01282-1 image captions).
Which to pick: For paraffin-section IHC, M01282-2 is the monoclonal option with documented EDTA retrieval and 1:50 staining; its captions do not report the fixative (catalog clone; M01282-2 IHC captions). For IF, A01282-1 has a rat-liver image, while M01282-2 lists both IF and ICC without an IF image in this catalog (A01282-1 IF caption; M01282-2 applications/images). For cross-species work, both list human, mouse and rat reactivity, but the supplied images provide no mouse staining example (catalog reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P61956 (SUMO2_HUMAN, Small ubiquitin-related modifier 2).
- Human Protein Atlas. SUMO2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SUMO2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear bodies. In addition localized to the calyx, connecting piece and flagellar centriole. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. SUMO2 antibody validation summary (3 antibodies).
- Exploration of SUMO2/3 Expression Levels and Autophagy Process in Fragile X-Associated Tremor/Ataxia Syndrome: Addressing Study Limitations and Insights for Future Research. Cells 2023 — PMC10571773.
- Novel proteomics and neuropathology of NOTCH2NLC-related neuronal intranuclear inclusion disease. Frontiers in aging neuroscience 2026 — PMC13375458.
- Small ubiquitin-related modifier 2/3 interacts with p65 and stabilizes it in the cytoplasm in HBV-associated hepatocellular carcinoma. BMC cancer 2015 — PMC4603762.
- Variation in expression of small ubiquitin-like modifiers in injured sciatic nerve of mice. Neural regeneration research 2019 — PMC6524499.
- PubMed PMID:8630065 — UniProt-cited evidence.
- PubMed PMID:9119407 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SUMO2 (Small ubiquitin-related modifier 2). Expected localisation General nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M01282-2). Validated IHC antibody: M01282-2 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular 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 lung cancer tissue; fixative not specified (datasheet M01282-2); 2. Fixation: Image fixative and duration unreported (datasheet M01282-2); 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 M01282-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M01282-2); 8. Primary antibody: Rabbit monoclonal (clone IBG-19) anti-SUMO2, 1:50 (datasheet M01282-2); 9. Primary incubation: Overnight at 4 °C (datasheet M01282-2). Watch for: Fixation: Keep fixation consistent across samples (standard IHC practice; not target-specific); Caveat: SUMO3 cross-detection may confound scoring (datasheet M01282-2); Isoform / epitope: 2 isoforms; residues 94–95 removed; epitope impact unknown (UniProt); Regulation: Regulation not specified (UniProt). Evidence: UniProt P61956, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm and nuclear bodies. In addition…, Datasheet M01282-2, PMC10571773, PMC6524499. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SUMO2/immunohistochemistry-ihc-guide.