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UBE2N / Ubiquitin-conjugating enzyme E2 N · IHC design guide
Design Immunohistochemistry for UBE2N
This guide covers cytoplasmic and nuclear UBE2N staining in paraffin sections, with the strongest reported signal in late spermatids (HPA tissue IHC). It includes an IHC starting concentration of 2–5 μg/mL for catalog antibody A02462-2 in human sections (datasheet A02462-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBE2N 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 | Cytoplasmic and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining; strongest in testis spermatids (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02462-2) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02462-2) | |
| Caveat | Assay may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation is unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms or processing; one 1–152 chain (UniProt) |
Section 1
Recommended UBE2N IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 heat retrieval (datasheet: A02462-2). Four published UBE2N IHC methods follow (PMC6639966; PMC9945078; PMC10039148; PMC11075218).
| Sample | Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A02462-2) |
| Fixation | Image fixative and duration unreported (datasheet A02462-2); 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 A02462-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02462-2) |
| Primary antibody | Rabbit anti-UBE2N, 2-5 μg/ml (datasheet A02462-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02462-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02462-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBE2N-positive staining in elongated or late spermatids of testis (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic nuclear expression, most abundant in testis. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A02462-2). Published alternatives use citrate buffer (PMC6639966) or Tris-EDTA pH 9.0 (PMC9945078).
Section 2
What Is the Expected UBE2N Staining Pattern?
UBE2N is found in the nucleus and cytoplasm (UniProt P61088: subcellular location), and HPA describes general nuclear and cytoplasmic tissue staining, most abundant in testis (HPA: tissue IHC profile). In paraffin sections, elongated or late spermatids are the clearest expected positive cells, with High staining (HPA: High in testis spermatids). HPA rates the tissue pattern Supported, while cautioning that its antibody targets proteins from more than one gene (HPA: reliability description).
| Nuclear and cytoplasmic staining in elongated or late spermatids, stronger than nearby weakly stained cells (HPA: High in testis spermatids; tissue IHC profile). | This fits the reported positive cell population and both annotated compartments (HPA: testis IHC; UniProt P61088: subcellular location). Judge the signal by identifiable cells and compartments, since HPA's Supported pattern includes a multiple-gene antibody caution (HPA: reliability description). |
| A sharply membrane-restricted pattern dominates otherwise intact cells (UniProt P61088: no transmembrane segment). | Treat this as unexpected and investigate localisation or staining artefact: UBE2N has no annotated transmembrane segment and is reported in the nucleus and cytoplasm (UniProt P61088: topology and subcellular location). A membrane outline alone does not establish UBE2N-positive cells. |
| Strong signal appears in adipocytes or lung alveolar type I cells (HPA: Not detected in these cell types). | These are cell-specific discrepancies, not proof that the whole tissue is negative (HPA: tissue IHC). Check cross-reactivity and, with chromogenic detection, endogenous enzyme activity as possible causes (general IHC practice). HPA's antibody caution makes an independent specificity check useful (HPA: reliability description). |
| Diffuse chromogen covers several compartments and extracellular areas without clear cell boundaries. | This cannot be scored as the reported nuclear and cytoplasmic pattern (HPA: tissue IHC profile; UniProt P61088: subcellular location). Background from detection or nonspecific binding can obscure the expected cell contrast (general IHC practice); compare a matched negative control before interpreting intensity. |
| Elongated or late spermatids lack convincing staining in an otherwise interpretable testis section (HPA: High in testis spermatids). | The result conflicts with HPA's clearest positive reference (HPA: testis IHC). First confirm that the relevant spermatids are present, then assess the staining run with a known-positive section and controls (general IHC practice). An absent signal alone cannot establish absent UBE2N. |
💡Expected UBE2N appearanceCall a section positive when identifiable elongated or late spermatids show High nuclear or cytoplasmic staining (HPA: High in testis spermatids; UniProt P61088: subcellular location); diffuse extracellular colour or a dominant membrane outline is an unexpected result, not that cell-specific pattern (HPA: tissue IHC profile; UniProt P61088: topology).
| Cell and tissue context (HPA: tissue IHC). | HPA reports Medium staining in adrenal, appendix, breast and duodenal glandular cells, bronchial respiratory epithelium, cerebral cortex neuropil and colon endothelium (HPA: tissue IHC). Its Not detected entries name particular cell populations; they do not define an entire organ as a negative control (HPA: tissue IHC). |
| Antibody evidence (HPA: reliability description; antibody validation). | The tissue profile is Supported and antibody HPA044976 is IHC Supported, with a warning that the antibody targets proteins from more than one gene (HPA: tissue reliability; antibody validation). Interpret unexpected staining with that specificity limit in mind; these labels do not establish that every positive cell contains UBE2N alone. |
| Topology and processing (UniProt P61088: topology and processing). | UBE2N is a 152-residue chain with no annotated transmembrane segment, signal peptide or propeptide (UniProt P61088: topology and processing). Those annotations support checking intracellular localisation; they do not specify antigen retrieval conditions or predict a fixation effect. |
| IF/ICC Q: What localisation should a separate IF/ICC guide assess? | A: HPA reports mainly nucleoplasmic staining and additional nucleoli fibrillar-center staining in ICC-IF, both Supported, with a multiple-gene antibody caution (HPA: subcellular ICC-IF). That cell-imaging observation can inform localisation review; it is not a paraffin IHC intensity standard (HPA: ICC-IF; tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in identifiable elongated or late spermatids (HPA: High in testis spermatids). | The expected positive population may be absent from the viewed area, or the staining run may have failed (HPA: testis IHC; general IHC practice). HPA supplies no target-specific fixation-sensitivity claim here. | Verify cell identity on the counterstain and examine a known-positive testis section in the same run (HPA: High in testis spermatids; general IHC practice). Review retrieval and detection using the validated IHC procedure, without inferring a UBE2N-specific fixation effect. |
| All compartments and empty spaces show diffuse chromogen. | Nonspecific binding, incomplete blocking, or detection background can hide cellular staining (general IHC practice). This appearance cannot be matched to HPA's cellular nuclear and cytoplasmic profile (HPA: tissue IHC profile). | Compare a matched negative control, inspect blocking and washing, and adjust the staining run according to the validated IHC procedure (general IHC practice). Score only resolvable cells and compartments against the HPA reference (HPA: tissue IHC). |
| Strong colour appears in adipocytes, skin fibroblasts, or lung alveolar type I cells (HPA: Not detected in those cell types). | The result could reflect cross-reactivity or endogenous chromogenic detection activity (general IHC practice). HPA also cautions that its antibody targets proteins from more than one gene (HPA: reliability description). | Confirm the stained cell identity, compare a no-primary control, and check the endogenous enzyme blocking step for the detection system (general IHC practice). Treat these HPA entries as cell-specific references, not whole-tissue exclusions (HPA: tissue IHC). |
| Staining forms a crisp cell-surface rim with little intracellular signal. | A membrane-dominant result conflicts with the annotated nuclear and cytoplasmic location and lack of a transmembrane segment (UniProt P61088: subcellular location and topology). It may reflect artefact or off-target staining. | Recheck morphology, compare the no-primary control, and review specificity with an independent control if available (general IHC practice). Do not count a membrane rim alone as the expected UBE2N pattern (UniProt P61088: subcellular location and topology). |
| Nuclear-only staining is seen where the paraffin-section pattern was expected to include cytoplasmic signal (HPA: tissue IHC profile). | HPA's ICC-IF summary is mainly nucleoplasmic, whereas its tissue IHC summary describes general nuclear and cytoplasmic expression (HPA: subcellular ICC-IF; tissue IHC). The methods provide different observations; this discrepancy alone does not identify a cause. | Record nuclear and cytoplasmic scores separately, check the relevant positive cell type, and compare like-for-like paraffin sections before calling the pattern discordant (HPA: tissue IHC; general IHC practice). Keep the ICC-IF localisation observation as context (HPA: subcellular ICC-IF). |
| Medium staining in a sampled tissue is mistaken for a failed positive result (HPA: Medium in several listed cell types). | The strongest supplied reference is testis elongated or late spermatids; HPA assigns Medium to several other specified populations (HPA: tissue IHC). Intensity expectations depend on the sampled cell population. | Identify and score the actual cell type, then compare it with its HPA entry: for example, bronchial respiratory epithelial cells or colon endothelial cells are Medium (HPA: tissue IHC). Use testis spermatids when a High reference is needed (HPA: testis IHC). |
Experiment evidence
IHC & IF images showing UBE2N staining
A02462-2 · IHCIHC analysis of Ubc13/UBE2N using anti-Ubc13/UBE2N antibody (A02462-2). Ubc13/UBE2N was detected in a paraffin-embedded section of human cer
A02462-2 · IHCIHC analysis of Ubc13/UBE2N using anti-Ubc13/UBE2N antibody (A02462-2). Ubc13/UBE2N was detected in a paraffin-embedded section of human bre
A02462-2 · IFIF analysis of Ubc13/UBE2N using anti-Ubc13/UBE2N antibody (A02462-2). Ubc13/UBE2N was detected in an immunocytochemical section of A549 cel
A02462-1 · IHCImmunohistochemistry of UBC13 in mouse thymus tissue with UBC13 antibody at 2 μg/mL.
A02462-1 · IHCImmunohistochemistry of UBC13 in mouse thymus tissue with UBC13 antibody at 2.5 μg/ml.
A02462-1 · IFImmunofluorescence of UBC13 in Mouse Thymus cells with UBC13 antibody at 10 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000177889-UBE2N/tissue/cerebral+cortex#imid_13823975
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000177889-UBE2N/tissue/stomach+1#imid_13822581
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000177889-UBE2N/tissue/colon#imid_13822607
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000177889-UBE2N/tissue/liver#imid_13822608
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA044976; green = UBE2N, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA044976; green = UBE2N, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBE2N IHC & IF
🧪Run testis first and expect elongated or late spermatids to stain (HPA: High in elongated or late spermatids). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); within the testis section, internal negative cells should show no specific chromogenic signal, but the supplied HPA rows do not identify a negative testicular cell type.
Positive control tissue: Testis (Elongated or late spermatids, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UBE2N in CACO-2, RT-4, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary or secondary-only slide, a concentration-matched rabbit isotype control, and a UBE2N knockout sample or validated peptide-block control where available (standard IHC practice; selected caption: rabbit primary antibody). For HRP/DAB staining of testis, quench endogenous peroxidase and assess background on the control slides (selected caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02462-2 paraffin-section caption does not state the fixative (selected caption: fixative not stated). The caption reports heat-mediated retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml, but does not establish that retrieval is required for every specimen (selected caption). Frozen-section ease is unreported; HPA shows ICC-IF images in CACO-2, RT-4 and U2OS with supported nucleoplasmic localisation, while a testis-specific artefact is not documented in the supplied evidence (HPA: subcellular data; HPA: High in elongated or late spermatids).
HPA tissue IHC evidence for UBE2N
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells type I | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skin | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBE2N IHC Tips
Troubleshoot UBE2N staining in paraffin section IHC by checking retrieval, compartment, controls and cell specific scoring before interpreting DAB signal.
What retrieval should I start with if UBE2N staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for this catalog antibody (datasheet A02462-2). The paraffin section example used that retrieval before 2 μg/ml primary antibody overnight at 4°C, so reproduce those documented conditions before changing several variables together (datasheet A02462-2). If staining remains weak, adjust heating duration in a small pilot while keeping the EDTA buffer and detection steps constant, and check tissue preservation after retrieval (standard IHC practice). Compare a positive control section in every run; the documented example is human cervical cancer tissue, although its fixative is unreported (datasheet A02462-2).
Could fixation explain weak or uneven UBE2N IHC staining?
Target specific sensitivity to fixation is unknown: the selected UBE2N caption identifies a paraffin section but does not state its fixative (datasheet A02462-2). Record the fixation method and duration for each specimen, then compare sections processed under matched conditions before attributing a staining difference to UBE2N abundance (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary antibody at the documented 2 μg/ml while evaluating those sections (datasheet A02462-2). Check morphology and internal staining consistency across the section; patchy loss associated with visibly poor preservation warrants repeating preparation or using a better preserved specimen (standard IHC practice).
Should UBE2N staining be nuclear, cytoplasmic or both?
Evaluate both nuclear and cytoplasmic staining in paraffin sections, because both compartments are annotated for UBE2N (UniProt P61088 subcellular location; HPA tissue IHC profile). Score them separately against the hematoxylin counterstain so dense nuclear DAB does not obscure a weaker cytoplasmic component (standard IHC practice). A predominantly nucleoplasmic pattern is also reported in cell imaging, with an additional nucleolar fibrillar center signal, but that evidence carries a multiple gene antibody caution (HPA subcellular). If staining is exclusively extracellular or confined to damaged edges, examine morphology and controls before calling it UBE2N; the protein has no transmembrane segment (UniProt P61088 topology; standard IHC practice).
How should epitope accessibility affect my interpretation of UBE2N IHC?
The supplied record lists 0 isoforms and a UBC core spanning residues 3–149, but it does not map this antibody’s epitope (UniProt P61088; datasheet A02462-2). Avoid claiming that retrieval exposes a particular residue or distinguishes an isoform when neither conclusion is supported (UniProt P61088; datasheet A02462-2). UBE2N has no signal peptide, propeptide or transmembrane segment, so interpret nuclear and cytoplasmic staining as intracellular signal rather than membrane sided staining (UniProt P61088 processing and topology). Its recorded acetylation at lysine 82 does not establish an effect on this antibody’s staining; compare controls under the same retrieval conditions (UniProt P61088 modified residues; standard IHC practice).
How can IF help assess a questionable UBE2N IHC pattern?
Use IF/ICC as a separate localisation check, not as a substitute for the documented paraffin section IHC conditions (datasheet A02462-2; standard IHC practice). Multiplex UBE2N with a marker identifying the expected cell type, such as a validated late spermatid marker when examining testis, because HPA reports high staining in elongated or late spermatids (HPA tissue IHC). Choose a far red fluorophore when shorter wavelength tissue autofluorescence interferes, and include single channel and no primary controls (standard IF practice). Because UBE2N has no transmembrane segment and is annotated in nucleus and cytoplasm, use controlled permeabilisation to access intracellular epitopes; validate its strength separately for IF (UniProt P61088 topology and subcellular location; standard IF practice).
How do I reduce diffuse or misleading DAB background?
First compare the section with a no primary control and inspect whether staining follows intact cells rather than folds, damaged edges or deposits (standard IHC practice). The documented paraffin section workflow used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and an anti rabbit secondary for 30 minutes at 37°C (datasheet A02462-2). A peroxidase block before DAB development helps distinguish endogenous enzyme activity from antibody dependent signal; treat that as a general chromogenic IHC step (standard IHC practice). If background persists, shorten DAB development or titrate the primary antibody while checking that the positive control retains nuclear or cytoplasmic staining (UniProt P61088 subcellular location; standard IHC practice).
How should I quantify UBE2N across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell classes and nuclear versus cytoplasmic compartments before scoring, since UBE2N is reported in both locations (UniProt P61088 subcellular location; HPA tissue IHC profile). For cell based comparisons, report the percentage of positive cells and an H score from 0–300, using consistent intensity bins and the same DAB threshold across slides (standard IHC practice). Normalise positive counts to the number of evaluable cells in each class, or report positive cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Exclude necrotic and folded areas, and compare batches with a matched control section because staining and development conditions affect measured intensity (standard IHC practice).
What distinguishes plausible UBE2N signal from an IHC artefact?
A plausible result follows intact cell morphology and can include nuclear or cytoplasmic staining, matching the annotated UBE2N compartments (UniProt P61088 subcellular location; HPA tissue IHC profile). In testis, high signal in elongated or late spermatids is consistent with HPA tissue IHC, whereas absence in adipocytes has also been reported (HPA tissue IHC). Treat isolated extracellular deposits, staining concentrated at section edges or necrosis, and DAB signal reproduced in a no primary control as artefact candidates (standard IHC practice). Check peroxidase blocking and repeat a questionable result with matched retrieval and controls; HPA’s supported tissue profile also cautions that its antibodies may target proteins from more than one gene (standard IHC practice; HPA tissue IHC reliability).
Boster reagents
Best UBE2N / Ubiquitin-conjugating enzyme E2 N IHC Antibodies
Two catalog antibodies have IHC images from human paraffin sections and mouse thymus tissue, plus IF images from A549 and mouse thymus cells (catalog image captions).
Real IHC data
Anti-Ubc13/UBE2N Antibody ®
Cat # A02462-2
Real IHC data
Anti-UBC13 UBE2N Antibody
Cat # A02462-1
A02462-2 will render with human cervical cancer paraffin-section IHC; its catalog also shows IF in A549 cells (A02462-2 image captions). A02462-1 will render with mouse thymus IHC; its catalog also shows IF in mouse thymus cells (A02462-1 image captions).
Which to pick: Choose A02462-2 for human paraffin-section IHC because its own caption documents EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (A02462-2 IHC caption). For IF/ICC, choose A02462-2 for A549 cells or A02462-1 for mouse thymus cells, according to their respective IF images (catalog IF captions). For work across species, both list human, mouse and rat reactivity, while the pictured IHC evidence is human tissue for A02462-2 and mouse thymus for A02462-1; the mouse tissue fixative is unreported (catalog reactivity; respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P61088 (UBE2N_HUMAN, Ubiquitin-conjugating enzyme E2 N).
- Human Protein Atlas. UBE2N tissue IHC expression (reliability: Supported).
- Human Protein Atlas. UBE2N subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli fibrillar center. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. UBE2N antibody validation summary (1 antibodies).
- Spatial UBE2N protein expression indicates genomic instability in colorectal cancers. BMC cancer 2019 — PMC6639966.
- Potential prognostic and predictive value of UBE2N, IMPDH1, DYNC1LI1 and HRASLS2 in colorectal cancer stool specimens. Biomedical reports 2023 — PMC9945078.
- SP1 transcriptionally regulates UBE2N expression to promote lung adenocarcinoma progression. Molecular biomedicine 2023 — PMC10039148.
- UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells. Biology direct 2024 — PMC11075218.
- PubMed PMID:8902611 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:16122702 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBE2N (Ubiquitin-conjugating enzyme E2 N). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02462-2). Validated IHC antibody: A02462-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Elongated or late spermatids); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human cervical cancer tissue; fixative not specified (datasheet A02462-2); 2. Fixation: Image fixative and duration unreported (datasheet A02462-2); 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 A02462-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02462-2); 8. Primary antibody: Rabbit anti-UBE2N, 2-5 μg/ml (datasheet A02462-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02462-2). Watch for: Fixation: Keep fixation consistent across sections (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 A02462-2); Caveat: Assay may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: No annotated isoforms or processing; one 1–152 chain (UniProt); Regulation: Expression regulation is unreported (UniProt). Evidence: UniProt P61088, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A02462-2, PMC6639966, PMC9945078. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBE2N/immunohistochemistry-ihc-guide.