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UBA2 / SUMO-activating enzyme subunit 2 · IHC design guide
Design Immunohistochemistry for UBA2
Plan paraffin IHC around nuclear staining in most cell types, with placental trophoblasts as an exception (HPA tissue IHC). The catalog antibody has a documented paraffin IHC workflow; score staining by cell population (datasheet A03816-2; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBA2 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 | Nuclear staining in tissue; cytoplasmic shuttling is possible (HPA tissue IHC; UniProt) | |
| Staining pattern | Nuclear staining in nearly all cell types except trophoblasts (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03816-2) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Placenta |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03816-2) | |
| Caveat | Placental trophoblasts may lack detectable staining (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt; datasheet A03816-2) |
Section 1
Recommended UBA2 IHC & IF Protocols
Compare the catalog antibody’s citrate pH 6 IHC-P protocol (datasheet A03816-2) with published colorectal, gastric, and synovial tissue protocols (PMC6236309; PMC6235804; PMC7526534).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A03816-2) |
| Fixation | Image fixative and duration unreported (datasheet A03816-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A03816-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03816-2) |
| Primary antibody | Rabbit anti-UBA2, 0.5-1μg/ml (datasheet A03816-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03816-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03816-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBA2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in essentially all cell types, except for placental trophoblasts. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet A03816-2). The colorectal protocol reports citrate steam retrieval at 120°C for 10 min (PMC6236309).
Section 2
What Is the Expected UBA2 Staining Pattern?
UBA2 should stain mainly in nuclei across essentially all cell types in paraffin-section IHC; placental trophoblasts are the reported exception (HPA: tissue IHC profile). Cytoplasmic signal can occur because UBA2 shuttles between nucleus and cytoplasm, but it should be interpreted against the predominantly nuclear tissue pattern (UniProt Q9UBT2: subcellular location; HPA: tissue IHC profile). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Clear nuclear staining in many cell types, including adipocytes, glandular cells or hematopoietic cells. | This matches the broad nuclear pattern. HPA reports High staining in these cell types in specified tissues; their intensity does not set a threshold for every section or preparation (HPA: tissue IHC profile and listed tissue observations). |
| Predominantly cytoplasmic staining with little or no nuclear signal. | Review localisation and background controls: UBA2 can shuttle through the cytoplasm, so cytoplasmic staining alone is not proof of artefact, but a cytoplasm-only result departs from HPA's predominantly nuclear IHC pattern (UniProt Q9UBT2: subcellular location; HPA: tissue IHC profile). |
| Strong staining assigned specifically to placental trophoblasts. | Investigate cell identification and nonspecific staining. HPA reports UBA2 as Not detected in trophoblastic cells; this observation does not establish that every other placental cell should be negative (HPA: placenta tissue IHC observation). |
| Diffuse chromogen across cells and extracellular areas, obscuring nuclei. | Treat the result as uninterpretable until background is controlled. Examine a primary-antibody omission control and the detection system for nonspecific or endogenous activity; those are general chromogenic IHC checks, not reported UBA2 properties (general IHC practice). |
| No staining in a section expected to contain HPA High cells. | Check whether the relevant cells are present, then review the IHC-validated antibody, retrieval and detection controls. An absent signal alone cannot distinguish low assay sensitivity from a biological difference (HPA: listed tissue observations; general IHC practice). |
💡Expected UBA2 appearanceCall a positive result when interpretable nuclear chromogen is seen in the relevant cells, potentially strong in HPA High cell types; broad diffuse background or convincing trophoblast staining warrants review (HPA: tissue IHC profile and listed tissue observations; general IHC practice).
| Cell type and tissue context | HPA describes nuclear expression in essentially all cell types, with trophoblasts excepted; High examples include adipocytes, adrenal glandular cells and bone marrow hematopoietic cells (HPA: tissue IHC profile and listed tissue observations). |
| Nuclear and cytoplasmic distribution | UniProt records both compartments and nuclear–cytoplasmic shuttling; HPA tissue IHC nevertheless describes a nuclear pattern. Score the dominant compartment and assess mixed staining in its cellular context (UniProt Q9UBT2: subcellular location; HPA: tissue IHC profile). |
| Isoforms and epitope coverage | Two isoforms are listed, but the supplied records do not locate an antibody epitope or establish isoform-specific staining. Verify the IHC-validated antibody's stated epitope before making an isoform claim (UniProt Q9UBT2: isoforms; supplied HPA antibody record). |
| Protein topology and processing | UBA2 has no transmembrane segment, signal peptide or propeptide, and its annotated chain spans residues 1–640. These annotations do not by themselves predict retrieval needs or fixation sensitivity (UniProt Q9UBT2: topology and processing). |
| IF/ICC Q&A: where should UBA2 appear? | Mainly in the nucleoplasm; HPA calls that location Supported in ICC-IF. Cytoplasmic localisation is also recorded by UniProt, so assess additional signal with controls and imaging context (HPA: subcellular ICC-IF; UniProt Q9UBT2: subcellular location). |
| Evidence strength | HPA rates tissue IHC Supported, describes medium agreement with RNA data, and lists antibody HPA041436 as Supported for IHC. Treat the pattern as a reference for interpretation, not a guaranteed intensity for each specimen (HPA: tissue IHC reliability and antibody record). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA High cells are present, but nuclei show no chromogen. | The assay may have insufficient signal, or the expected cell population may have been misidentified; this result does not establish a UBA2-specific retrieval problem (HPA: listed tissue observations; general IHC practice). | Confirm the cell population, check a known-positive section and detection controls, then review the IHC-validated antibody's IHC conditions (HPA: listed tissue observations; general IHC practice). |
| Signal appears cytoplasmic with weak or absent nuclear staining. | UBA2 can shuttle, but a cytoplasm-dominant result conflicts with the principal HPA tissue pattern and may include background (UniProt Q9UBT2: subcellular location; HPA: tissue IHC profile). | Reassess nuclear boundaries with the counterstain; compare a known-positive tissue and an omission control before interpreting cytoplasmic signal (general IHC practice). |
| Placental trophoblasts appear strongly positive. | This differs from HPA's Not detected trophoblast observation; cell assignment or nonspecific chromogen may explain the discrepancy (HPA: placenta tissue IHC observation; general IHC practice). | Verify trophoblast identity on the section and compare primary-antibody omission and positive-tissue controls; report a reproducible discrepancy explicitly (general IHC practice). |
| Staining is diffuse or extracellular and nuclei cannot be scored. | Nonspecific antibody or detection signal, including endogenous detection activity, can obscure cellular localisation in chromogenic IHC (general IHC practice). | Inspect omission and detection-only controls; adjust blocking, washing or detection conditions according to the assay controls before scoring (general IHC practice). |
| Signal intensity varies among cell types in one section. | HPA reports High staining in selected cell populations but does not provide a universal intensity cutoff; heterogeneous staining needs cell-specific interpretation (HPA: listed tissue observations). | Score identifiable cell populations separately, record nuclear localisation and compare with appropriate controls rather than assigning one score to the whole section (HPA: tissue IHC profile; general IHC practice). |
| ICC-IF shows signal outside the nucleoplasm. | HPA's main ICC-IF location is the nucleoplasm, while UniProt also records cytoplasmic UBA2 and shuttling; imaging background remains possible (HPA: subcellular ICC-IF; UniProt Q9UBT2: subcellular location). | Check nuclear segmentation and fluorescence controls, then describe any reproducible cytoplasmic component alongside the dominant compartment (general IF practice). |
Experiment evidence
IHC & IF images showing UBA2 staining
A03816-2 · IHCIHC analysis of SAE2/UBA2 using anti-SAE2/UBA2 antibody (A03816-2). SAE2/UBA2 was detected in paraffin-embedded section of human colon cance
A03816-2 · IHCIHC analysis of SAE2/UBA2 using anti-SAE2/UBA2 antibody (A03816-2). SAE2/UBA2 was detected in paraffin-embedded section of human mammary can
A03816-2 · IFIF analysis of SAE2/UBA2 using anti-SAE2/UBA2 antibody (A03816-2). SAE2/UBA2 was detected in immunocytochemical section of A431 cell. Enzyme
M03816-2 · IHCIHC analysis of SAE2/UBA2 using anti-SAE2/UBA2 antibody (M03816-2). SAE2/UBA2 was detected in paraffin-embedded section of human intestinal
M03816-2 · IHCIHC analysis of SAE2/UBA2 using anti-SAE2/UBA2 antibody (M03816-2). SAE2/UBA2 was detected in paraffin-embedded section of human intestinal
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000126261-UBA2/tissue/cerebral+cortex#imid_10756259
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000126261-UBA2/tissue/colon#imid_10756177
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000126261-UBA2/tissue/liver#imid_10756170
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000126261-UBA2/tissue/kidney#imid_10756171
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041436; green = UBA2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041436; green = UBA2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBA2 IHC & IF
🧪Run adipose tissue first and assess staining in adipocytes (HPA: High in adipocytes), then run placenta as the negative tissue and assess trophoblastic cells (HPA: Not detected in trophoblastic cells). On the adipose slide, treat nonadipocyte cells as candidate internal negatives only if independently shown to lack UBA2; they should then lack specific staining (standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Placenta (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UBA2 in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a concentration-matched nonimmune rabbit IgG isotype control for the rabbit primary, and UBA2-knockout tissue if available (caption: rabbit primary; standard IHC practice). For chromogenic detection, quench endogenous peroxidase and check endogenous biotin because the caption uses a biotin-based detection system (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (caption: fixative unreported). Citrate retrieval at pH 6 for 20 minutes is a documented starting condition, but retrieval dependency is unreported (caption: citrate retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; adipose processing may leave lipid vacuoles that complicate cell identification (standard histology practice).
HPA tissue IHC evidence for UBA2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Placenta | Trophoblastic cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBA2 IHC Tips
Troubleshoot UBA2 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, reference staining patterns, and controlled comparisons.
What retrieval conditions should I start with for weak UBA2 staining?
Start with heat-mediated antigen retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A03816-2). The selected paraffin-section image used this retrieval before overnight incubation with 1 μg/ml primary antibody at 4°C (catalog antibody A03816-2 tissue-IHC caption). If nuclear signal is weak, compare a modestly longer heating interval with the documented condition on adjacent sections while keeping detection constant (standard IHC practice). Check tissue morphology and background alongside signal, since excessive retrieval can impair interpretation (standard IHC practice). Record retrieval time and heating method so staining differences can be traced across runs (standard IHC practice).
How should I investigate fixation-related loss of UBA2 staining?
Target-specific fixation sensitivity for UBA2 is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (catalog antibody A03816-2 tissue-IHC caption). Record the fixative, fixation duration, and processing history for each specimen before comparing staining (standard IHC practice). Run sections with different known fixation histories through the same pH 6 citrate retrieval and detection conditions to assess a possible processing effect (datasheet A03816-2; standard IHC practice). Compare nuclear signal with morphology and a suitable tissue control on the same run (standard IHC practice). Do not assign weak staining to fixation without that controlled comparison (standard IHC practice).
Should UBA2 staining appear in nuclei, cytoplasm, or both?
Expect predominantly nuclear staining in paraffin sections: tissue IHC reports nuclear expression in essentially all cell types except placental trophoblasts (HPA: tissue IHC profile). Nucleoplasm is the supported main subcellular location in imaging data (HPA: subcellular summary). UBA2 can also occupy the cytoplasm and shuttle between cytoplasm and nucleus (UniProt Q9UBT2: subcellular location). Score nuclear and cytoplasmic chromogenic signal separately, using a counterstain to define nuclei (standard IHC practice). If staining appears exclusively cytoplasmic or follows tissue edges, compare it with a reagent control and the expected nuclear pattern before interpreting it as redistribution (HPA: tissue IHC profile; standard IHC practice).
Can this antibody distinguish UBA2 isoforms or modified epitopes in tissue?
UBA2 has 2 annotated isoforms, but the supplied antibody caption gives no epitope sequence or isoform specificity (UniProt Q9UBT2: isoforms; catalog antibody A03816-2 tissue-IHC caption). Therefore, chromogenic staining alone cannot assign signal to either isoform (standard IHC interpretation). The record lists phosphorylation and acetylation sites, including phosphoserines at residues 207, 507, and 592 (UniProt Q9UBT2: modified residues). Do not interpret staining intensity as modification state without a modification-specific reagent and appropriate controls (standard IHC practice). If isoform attribution matters, verify the antibody epitope and compare it with isoform sequences before designing tissue controls (standard IHC practice).
How can companion IF clarify an ambiguous chromogenic UBA2 pattern?
Use IF as a companion localisation check while keeping the paraffin-section chromogenic result as the IHC readout (standard IHC/IF practice). For multiplexing, pair UBA2 with a validated glandular-cell marker in appendix, where glandular cells have high tissue-IHC staining (HPA: High in appendix glandular cells; standard IF practice). Choose spectrally separated fluorophores after checking tissue autofluorescence, and include single-label controls for bleed-through (standard IF practice). UBA2 is mainly nucleoplasmic and has no transmembrane segment, so use permeabilisation appropriate for access to an intracellular nuclear epitope and titrate it against morphology (HPA: subcellular summary; UniProt Q9UBT2: topology; standard IF practice).
How do I distinguish UBA2 signal from chromogenic background?
The selected paraffin-section image used 10% goat serum blocking, a biotinylated secondary antibody, an avidin–biotin detection complex, and DAB (catalog antibody A03816-2 tissue-IHC caption). Include a primary-omission control to assess detection-system staining, and apply a peroxidase block when using DAB chemistry (standard IHC practice). If diffuse brown signal persists, compare blocking, washing, and primary-antibody concentration on adjacent sections while holding exposure conditions constant (standard IHC practice). Inspect areas with damaged morphology separately from intact cells before scoring (standard IHC practice). A convincing UBA2 pattern should retain interpretable nuclear signal relative to controls and the reported tissue distribution (HPA: tissue IHC profile; standard IHC practice).
What should I measure when comparing UBA2 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score the percentage of positive nuclei and nuclear staining intensity in defined cell populations; an H-score combines those measures on a consistent intensity scale (standard IHC practice). Report cytoplasmic staining separately because UBA2 can shuttle between nucleus and cytoplasm (UniProt Q9UBT2: subcellular location). Normalise positive-cell counts to all eligible cells of the same type within the assessed area, or report density per mm² of viable tissue (standard IHC practice). Exclude folds, edges, and necrotic regions using the same rules for every section (standard IHC practice). Keep retrieval, chromogen development, imaging, and scoring thresholds consistent across specimens (standard IHC practice).
When should I doubt an apparent UBA2-positive result?
Treat a nuclear pattern in intact cells as more consistent with the reported tissue profile than staining confined to damaged edges or necrotic areas (HPA: tissue IHC profile; standard IHC interpretation). Check cell identity before comparing intensity: placental trophoblasts were not detected in the tissue-IHC dataset, whereas appendix glandular cells were high (HPA: tissue IHC profile). Cytoplasmic signal is biologically plausible because UBA2 shuttles, but isolated cytoplasmic DAB without corroborating nuclear staining warrants control review (UniProt Q9UBT2: subcellular location; HPA: tissue IHC profile). Use primary-omission and peroxidase-block controls to assess detection artefacts and endogenous enzyme activity (standard IHC practice). Interpret any remaining difference alongside morphology and matched processing conditions (standard IHC practice).
Boster reagents
Best UBA2 / SUMO-activating enzyme subunit 2 IHC Antibodies
Catalog images show UBA2 IHC in paraffin sections of human cancers and mouse and rat testis, plus IF in A431 cells (catalog image captions).
Real IHC data
Anti-SAE2/UBA2 Antibody ®
Cat # A03816-2
Real IHC data
Anti-SAE2/UBA2 Antibody ® (monoclonal, 5B13)
Cat # M03816-2
A03816-2 has IHC images from human colon and mammary cancers and mouse and rat testis, plus an IF image from A431 cells (A03816-2 image captions). M03816-2 has IHC images from human intestinal cancer paraffin sections (M03816-2 image captions).
Which to pick: For tissue IHC across human, mouse and rat, choose A03816-2 because its paraffin-section images cover all three species (A03816-2 IHC captions). For human tissue IHC with a monoclonal antibody, choose M03816-2, clone 5B13; its supplied IHC images show human intestinal cancer (catalog clone; M03816-2 IHC captions). For IF/ICC, choose A03816-2, which has an A431 IF image; the fixative is unreported in both SKUs’ IHC captions (A03816-2 IF caption; A03816-2 and M03816-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UBT2 (SAE2_HUMAN, SUMO-activating enzyme subunit 2).
- Human Protein Atlas. UBA2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. UBA2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. UBA2 antibody validation summary (1 antibodies).
- UBA2 promotes proliferation of colorectal cancer. Molecular medicine reports 2018 — PMC6236309.
- Ubiquitin-like modifier activating enzyme 2 promotes cell migration and invasion through Wnt/β-catenin signaling in gastric cancer. World journal of gastroenterology 2018 — PMC6235804.
- Increased SUMO-activating enzyme SAE1/UBA2 promotes glycolysis and pathogenic behavior of rheumatoid fibroblast-like synoviocytes. JCI insight 2020 — PMC7526534.
- Knockdown of ubiquitin-like modifier-activating enzyme 2 promotes apoptosis of clear cell renal cell carcinoma cells. Cell death & disease 2021 — PMC8578554.
- PubMed PMID:9920803 — UniProt-cited evidence.
- PubMed PMID:10217437 — UniProt-cited evidence.
- PubMed PMID:10187858 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBA2 (SUMO-activating enzyme subunit 2). Expected localisation Nuclear staining in tissue; cytoplasmic shuttling is possible (HPA tissue IHC; UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03816-2). Validated IHC antibody: A03816-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Placenta. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A03816-2); 2. Fixation: Image fixative and duration unreported (datasheet A03816-2); 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: Citrate pH 6 , 20 min (datasheet A03816-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03816-2); 8. Primary antibody: Rabbit anti-UBA2, 0.5-1μg/ml (datasheet A03816-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03816-2). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03816-2); Caveat: Placental trophoblasts may lack detectable staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt; datasheet A03816-2); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q9UBT2, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A03816-2, PMC6236309, PMC6235804. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBA2/immunohistochemistry-ihc-guide.