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UBR2 / E3 ubiquitin-protein ligase UBR2 · IHC design guide
Design Immunohistochemistry for UBR2
Plan chromogenic UBR2 IHC on paraffin sections with the catalog antibody at 0.5–1 µg/mL (datasheet A05812-2). Compare its annotated nuclear location (UniProt) with the cytoplasmic and nuclear staining reported in tissues (HPA tissue IHC), and score each compartment separately.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBR2 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 staining (HPA tissue IHC) | |
| Staining pattern | Broad cytoplasmic and nuclear staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05812-2) | |
| 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 fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Cytoplasmic staining conflicts with nuclear annotation (HPA tissue IHC; UniProt) | |
| Regulation | DUSP22 promotes UBR2 degradation (UniProt) | |
| Isoform / epitope | 4 isoforms; verify epitope coverage (UniProt) |
Section 1
Recommended UBR2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A05812-2) with the published mouse TNBC protocol (PMC12271019).
| Sample | Paraffin-embedded human pancreatic cancer tissue; fixative not specified (datasheet A05812-2) |
| Fixation | Image fixative and duration unreported (datasheet A05812-2); 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 A05812-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05812-2) |
| Primary antibody | Rabbit anti-UBR2, 0.5-1μg/ml (datasheet A05812-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05812-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05812-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBR2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteTry EDTA pH 8.0 heat retrieval first: it matches both the catalog datasheet (A05812-2) and the published protocol (PMC12271019).
Section 2
What Is the Expected UBR2 Staining Pattern?
UBR2 is annotated in the nucleus and on meiotic chromosomes; it has no transmembrane segment (UniProt Q8IWV8). Tissue IHC reports widespread cytoplasmic and nuclear staining, including high staining in adipocytes and thyroid glandular cells (HPA tissue IHC). HPA rates its tissue staining Approved, with medium agreement with RNA data and a subcellular pattern contradicted by external data (HPA tissue IHC).
| Cell-associated nuclear staining, with some cytoplasmic staining, across several tissue compartments. | This fits the broad HPA tissue IHC profile. Nuclear signal agrees with UniProt localization; cytoplasmic signal is reported by HPA but has a subcellular evidence caveat. Assess staining within recognizable cells and compare compartments on the same section (HPA tissue IHC; UniProt Q8IWV8). |
| Strong staining in adipocytes or thyroid glandular cells, alongside weaker staining elsewhere. | High staining in these cell types is reported by HPA. Contrast can help orient interpretation, but HPA also reports widespread expression; a weaker neighboring population is not automatically a negative control (HPA tissue IHC). |
| Color is concentrated in acellular spaces or surface deposits, without a convincing intracellular pattern. | That distribution does not match the cell-associated tissue profile or nuclear localization. Treat it as a possible staining artefact and inspect a matched negative control; membrane signal alone needs caution because HPA ICC-IF also reports an additional plasma-membrane location (HPA tissue IHC; HPA ICC-IF; UniProt Q8IWV8; general IHC practice). |
| A single unexpected cell population stains strongly while expected high-staining cells do not. | Consider antibody cross-reactivity or endogenous detection activity before assigning that population as UBR2-positive. HPA lists high staining in defined cell types, but its low-staining entries are not negatives; compare morphology and controls before interpreting the difference (HPA tissue IHC; general IHC practice). |
| Weak, diffuse color covers cells and surrounding tissue, or a known high-staining tissue has no signal. | Diffuse color limits compartment scoring and suggests background. Absent signal in an expected high-staining population calls for a run-level check of controls and detection steps; neither observation alone establishes UBR2 absence (HPA tissue IHC; general IHC practice). |
💡Expected UBR2 appearanceA credible positive has cell-associated nuclear staining, potentially with cytoplasmic staining, in broadly distributed cells; adipocytes and thyroid glandular cells may stain strongly, while diffuse acellular color is suspect (UniProt Q8IWV8; HPA tissue IHC; general IHC practice).
| Compartment evidence | UniProt places UBR2 in the nucleus and associates it with meiotic chromatin; HPA tissue IHC reports nuclear and cytoplasmic staining and flags external disagreement about subcellular location. Score compartments separately rather than treating all cytoplasmic color as confirmed localization (UniProt Q8IWV8; HPA tissue IHC). |
| Tissue comparison | HPA reports high staining in adipocytes, adrenal and thyroid glandular cells, and smooth muscle cells; pancreatic exocrine glandular cells are listed as low. These are observed staining levels, not guaranteed outcomes for every section or an absolute positive/negative tissue pair (HPA tissue IHC). |
| Antibody evidence | HPA027869 and HPA027880 each have Approved IHC status. HPA's overall tissue rating is Approved with medium RNA–staining consistency; these labels support cautious comparison with its reported pattern, not perfect specificity in a new preparation (HPA antibodies; HPA tissue IHC). |
| Protein forms and topology | UniProt lists 4 isoforms, no signal peptide or propeptide, and no transmembrane segment. The payload gives no antibody epitope or isoform coverage, so a compartment difference cannot be assigned to a particular isoform or shedding event (UniProt Q8IWV8). |
| What should IF/ICC show? | HPA ICC-IF reports mainly nucleoplasmic signal and an additional plasma-membrane location, both Approved, with images in A-431, U-251MG and U2OS cells. Use that as a localization cross-check; its cell-image pattern does not resolve the tissue IHC subcellular disagreement (HPA ICC-IF; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining cells are blank. | The section or detection run may have failed; absence in one slide cannot establish absent UBR2 (HPA tissue IHC; general IHC practice). | Check a same-run positive tissue and detection controls, then review primary-antibody incubation and chromogenic development. If the run still fails, optimize retrieval as a general IHC step; UBR2-specific fixation or retrieval sensitivity is unreported here (general IHC practice). |
| The entire section has diffuse brown color. | Non-specific background or endogenous detection activity may obscure cellular staining (general IHC practice). | Compare a no-primary control and inspect blocking, washes and chromogen exposure. Interpret UBR2 only where cell boundaries and compartments remain distinguishable (general IHC practice). |
| Strong color appears only in an unexpected cell population. | Cross-reactivity or endogenous activity is possible, especially if HPA high-staining cell types are blank; HPA low-staining cells are not defined negatives (HPA tissue IHC; general IHC practice). | Compare the no-primary control, cell morphology and a high-staining reference tissue in the same run. Record the unexpected distribution rather than calling it specific from intensity alone (HPA tissue IHC; general IHC practice). |
| Staining is mainly cytoplasmic, with little nuclear color. | HPA tissue IHC reports cytoplasmic and nuclear expression, while UniProt annotates nuclear localization; HPA flags external subcellular disagreement (HPA tissue IHC; UniProt Q8IWV8). | Score nuclear and cytoplasmic signals separately and compare a control section and the reported HPA pattern. Avoid assigning the cytoplasmic component to a UBR2 isoform without epitope evidence (HPA tissue IHC; UniProt Q8IWV8; general IHC practice). |
| Color pools in lumina or outside intact cells. | Acellular deposition does not match the reported cellular tissue pattern (HPA tissue IHC; general IHC practice). | Inspect the section and no-primary control for deposits or residual chromogen. Exclude acellular color from cell-level scoring (general IHC practice). |
| Pancreatic exocrine cells stain weakly while another tissue stains strongly. | HPA lists pancreatic exocrine glandular cells as low despite UniProt describing high overall pancreatic expression and protein in acinar cells; the sources measure or describe different aspects of expression (HPA tissue IHC; UniProt Q8IWV8). | Do not use pancreatic exocrine cells as a required high-intensity IHC control. Compare with an HPA high-staining cell type and document the tissue and cellular compartment scored (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing UBR2 staining
A05812-2 · IHCIHC analysis of UBR2 using anti-UBR2 antibody (A05812-2). UBR2 was detected in paraffin-embedded section of human pancreatic cancer tissue.
A05812-2 · IHCIHC analysis of UBR2 using anti-UBR2 antibody (A05812-2). UBR2 was detected in paraffin-embedded section of human pancreatic cancer tissue.
A05812-2 · IFIF analysis of UBR2 using anti-UBR2 antibody (A05812-2). UBR2 was detected in immunocytochemical section of HEPG2 cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000024048-UBR2/tissue/cerebral+cortex#imid_7552920
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000024048-UBR2/tissue/colon#imid_7552854
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000024048-UBR2/tissue/liver#imid_7552847
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000024048-UBR2/tissue/kidney#imid_7552856
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027880; green = UBR2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027880; green = UBR2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBR2 IHC & IF
🧪Start with adipose tissue: adipocytes should stain (HPA: High in adipocytes). HPA detects UBR2 in all 45 scored tissues, so use no-primary and isotype controls as the negative references; no cell type on the positive slide is established as UBR2-negative, and any unstained neighboring cells are only provisional internal contrasts (HPA: no negative tissue rows).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: UBR2 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 UBR2 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Run a secondary-only section and a matched rabbit IgG isotype control of the same monoclonal or polyclonal format, plus UBR2-knockout material if available (caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase and check endogenous biotin when using the caption’s biotin-based DAB detection system (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: Paraffin-section IHC is demonstrated with heat retrieval in EDTA at pH 8.0, but the selected-SKU caption does not report the fixative or establish a target-specific fixation window or fixation effect (caption: A05812-2). Retrieval is part of that demonstrated procedure; its necessity relative to other conditions is unreported (caption: EDTA heat retrieval). Frozen sections and IF cannot be judged easier from the supplied evidence; in adipose tissue, lipid vacuoles leave little cell material to score, so assess cellular staining against the negative controls (HPA: High in adipocytes; standard tissue histology).
HPA tissue IHC evidence for UBR2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Subcellular location is contradicted by external 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 → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: UBR2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced UBR2 IHC Tips
Troubleshoot UBR2 staining in paraffin sections using the catalog antibody’s IHC evidence, with IF/ICC considerations kept to a single entry.
What retrieval conditions should I try first when UBR2 staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections stained with A05812-2 (datasheet A05812-2). The selected pancreatic cancer image used that retrieval before incubation with 1 μg/mL antibody overnight at 4°C (datasheet A05812-2). If signal is weak, compare retrieval heating times on matched sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Include a positive control section in each run so a failed retrieval step is distinguishable from limited UBR2 expression (standard IHC practice). Assess nuclear and cytoplasmic staining separately because both appear in tissue IHC (HPA: ubiquitous cytoplasmic and nuclear expression).
Could fixation explain variable UBR2 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown; the selected paraffin-section caption does not report its fixative (datasheet A05812-2). Record each block’s fixative, fixation duration and processing history before comparing staining across specimens (standard IHC practice). Compare matched sections using the same EDTA pH 8.0 retrieval and the same detection run to limit procedural variation (datasheet A05812-2; standard IHC practice). If one block stains poorly, check tissue preservation and a positive control before changing antibody concentration (standard IHC practice). Do not assign a UBR2-specific fixation effect from its nuclear location or phosphorylation sites; those observations do not establish fixation sensitivity (UniProt Q8IWV8).
How should I assess nuclear, cytoplasmic and membrane staining for UBR2?
Score nuclear and cytoplasmic staining separately because tissue IHC reports both compartments (HPA: ubiquitous cytoplasmic and nuclear expression). Nucleoplasm is the main approved IF location, with additional plasma-membrane staining, whereas UniProt lists nucleus and chromosome (HPA: subcellular location; UniProt Q8IWV8). UBR2 has no annotated transmembrane segment, so membrane-associated staining alone does not establish a membrane-spanning form (UniProt Q8IWV8 topology). Compare each compartment with matched positive and negative controls, and inspect whether staining follows intact cell boundaries rather than section edges (standard IHC practice). Treat an isolated membrane pattern cautiously because HPA describes a conflict between tissue staining location and external data (HPA: reliability description).
Can different UBR2 isoforms or epitope masking change the IHC pattern?
UBR2 has 4 listed isoforms, but the supplied antibody caption does not map its epitope or establish which isoforms it detects (UniProt Q8IWV8; datasheet A05812-2). Obtain the immunogen or epitope coordinates before interpreting a missing signal as loss of every isoform (standard antibody-validation practice). The record lists modified residues including phosphoserine 476 and 1694, yet it provides no evidence that these modifications alter this antibody’s binding (UniProt Q8IWV8). Compare matched sections under the documented EDTA pH 8.0 retrieval first, then evaluate any alternative retrieval condition as an optimisation experiment (datasheet A05812-2; standard IHC practice). Interpret compartment differences only after checking antibody specificity and morphology (standard IHC practice).
How can IF help check an ambiguous chromogenic UBR2 pattern?
Use IF/ICC as a separate cross-check of compartment assignment; the catalog caption documents paraffin-section chromogenic IHC, not an IF procedure (datasheet A05812-2). In pancreas, multiplex UBR2 with an acinar-cell marker to identify the cells of interest, since acinar-cell UBR2 is reported at protein level (UniProt Q8IWV8). Choose spectrally separated, preferably red or far-red fluorophores after inspecting unstained tissue autofluorescence (standard IF practice). For a nuclear epitope, test gentle permeabilisation; for a proposed membrane-associated epitope, assess detergent effects because UBR2 has no annotated transmembrane segment or established epitope orientation (UniProt Q8IWV8 topology; standard IF practice). Compare nuclear calls with the reported nucleoplasmic IF location (HPA: subcellular location).
How do I troubleshoot diffuse brown staining in UBR2 IHC?
Start by comparing a no-primary control with the stained section to identify signal from the detection system (standard IHC practice). The selected image used 10% goat serum, biotinylated secondary antibody, a streptavidin–biotin complex and DAB (datasheet A05812-2). With that workflow, check endogenous peroxidase blocking and possible endogenous biotin staining before attributing diffuse brown colour to UBR2 (standard IHC practice). Also inspect washing, reagent concentration and tissue folds, which can produce spatially uneven background (standard IHC practice). Evaluate signal against intact nuclear and cytoplasmic morphology because both compartments have been reported in tissue IHC (HPA: ubiquitous cytoplasmic and nuclear expression).
What is a defensible way to quantify UBR2 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysed cell population and compartment before scoring, since tissue IHC reports both nuclear and cytoplasmic UBR2 (HPA: ubiquitous cytoplasmic and nuclear expression). For each compartment, record percentage-positive cells and intensity grades 0–3, then calculate an H-score from 0–300 if graded intensity is reproducible (standard IHC scoring practice). Alternatively, report positive-cell density per mm² of viable tissue when cell counts are more reliable than intensity (standard IHC scoring practice). Normalise to the number of eligible cells or measured viable area, and apply one threshold across all comparison sections (standard IHC scoring practice). Exclude necrosis, folds and section edges from the scored region (standard IHC scoring practice).
When is a brown UBR2 signal convincing rather than artefactual?
A convincing call has staining within intact cells, a reproducible compartment pattern and a clean no-primary control (standard IHC practice). Nuclear staining is biologically plausible because UBR2 is listed in the nucleus and on chromosomes; cytoplasmic tissue staining is also reported (UniProt Q8IWV8; HPA: tissue IHC profile). Check cell identity rather than assuming every pancreatic compartment stains equally: acinar-cell protein expression is reported, while HPA scores exocrine glandular cells low (UniProt Q8IWV8; HPA: pancreas). Discount staining concentrated at edges, folds or necrotic zones, and investigate diffuse DAB signal from endogenous enzyme activity (standard IHC practice). Treat membrane-only staining cautiously in light of the reported localisation disagreement (HPA: reliability description).
Boster reagents
Best UBR2 / E3 ubiquitin-protein ligase UBR2 IHC Antibodies
A05812-2 has paraffin-section IHC images from human pancreatic and renal cancer tissue and an IF image from HEPG2 cells (A05812-2 IHC/IF image captions). Human and mouse reactivity is listed (A05812-2 catalog reactivity).
Real IHC data
Anti-UBR2 ® Antibody
Cat # A05812-2
A05812-2 will render with its human pancreatic cancer paraffin-section IHC figure; its caption documents EDTA retrieval and DAB detection (A05812-2 IHC image caption). The catalog also lists IF/ICC, and its IF caption documents staining in HEPG2 cells (A05812-2 application list; A05812-2 IF image caption).
Which to pick: Choose A05812-2 for chromogenic IHC on paraffin sections; its own figure documents that preparation, but does not report the fixative (A05812-2 IHC image caption). For IF/ICC, the same SKU has a cell-staining example (A05812-2 IF image caption). For human or mouse samples, A05812-2 lists both species as reactive, while the supplied images document human samples only (A05812-2 catalog reactivity; A05812-2 IHC/IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IWV8 (UBR2_HUMAN, E3 ubiquitin-protein ligase UBR2).
- Human Protein Atlas. UBR2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. UBR2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane..
- Human Protein Atlas. UBR2 antibody validation summary (2 antibodies).
- The role of UBR2 in triple-negative breast cancer and its implications for immune checkpoint blockade therapy. Discover oncology 2025 — PMC12271019.
- PubMed PMID:15548684 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBR2 (E3 ubiquitin-protein ligase UBR2). Expected localisation Cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05812-2). Validated IHC antibody: A05812-2 at 0.5-1μg/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: Paraffin-embedded human pancreatic cancer tissue; fixative not specified (datasheet A05812-2); 2. Fixation: Image fixative and duration unreported (datasheet A05812-2); 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 A05812-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05812-2); 8. Primary antibody: Rabbit anti-UBR2, 0.5-1μg/ml (datasheet A05812-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05812-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Cytoplasmic staining conflicts with nuclear annotation (HPA tissue IHC; UniProt); Isoform / epitope: 4 isoforms; verify epitope coverage (UniProt); Regulation: DUSP22 promotes UBR2 degradation (UniProt). Evidence: UniProt Q8IWV8, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05812-2, PMC12271019. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBR2/immunohistochemistry-ihc-guide.