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UBE2I / SUMO-conjugating enzyme UBC9 · IHC design guide
Design Immunohistochemistry for UBE2I
Plan chromogenic paraffin IHC for UBE2I using the catalog antibody at 0.5–1 μg/mL (datasheet A02295). Compare nuclear and cytoplasmic staining across cell types, with high staining in tonsil non-germinal center cells and undetected staining in adipocytes as tissue controls (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBE2I 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 and cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Most tissues show nuclear and cytoplasmic cell staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02295) | |
| Positive control | Bronchus+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 paraffin sections and controls. (selected-SKU IHC image A02295) | |
| Caveat | Staining has only medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No specific expression regulator is reported (UniProt) | |
| Isoform / epitope | No isoforms listed; the mature chain spans residues 2–158 (UniProt) |
Section 1
Recommended UBE2I IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A02295) is paired with a published ccRCC and adjacent kidney IHC protocol (PMC8861443 methods).
| Sample | Paraffin-embedded human appendicitis tissue; fixative not specified (datasheet A02295) |
| Fixation | Image fixative and duration unreported (datasheet A02295); 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 A02295) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02295) |
| Primary antibody | Rabbit anti-UBE2I, 0.5-1μg/ml (datasheet A02295) |
| Primary incubation | Overnight at 4 °C (datasheet A02295) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02295) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBE2I-positive staining in basal cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A02295), then compare the published primary dilution and incubation conditions (PMC8861443 methods).
Section 2
What Is the Expected UBE2I Staining Pattern?
UBE2I is mainly nuclear, with cytoplasmic and perinuclear localization also reported (UniProt P63279: subcellular location). In tissue IHC, expect nuclear and cytoplasmic staining in many cell types, including bronchial basal cells and placental trophoblastic cells with High staining (HPA: tissue IHC). UBE2I has no transmembrane segment, so a membrane-only pattern is unexpected (UniProt P63279: topology). HPA rates its tissue profile Approved, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Bronchial basal cells or placental trophoblastic cells stain strongly, with nuclear and cytoplasmic signal. | This fits HPA's High staining in those cell types and its nuclear and cytoplasmic tissue profile (HPA: tissue IHC). Score the named cells and compartments separately; a strong field-wide signal alone does not show that the expected cells stained. |
| Signal outlines cell membranes, with little nuclear or cytoplasmic staining. | A membrane-only pattern conflicts with the reported intracellular locations and lack of a transmembrane segment (UniProt P63279: subcellular location and topology). Treat it as suspect and compare it with a known-positive section stained in the same run. |
| Adipocytes stain strongly while expected positive cells remain faint. | HPA reports UBE2I as Not detected in adipocytes but High in bronchial basal cells and placental trophoblastic cells (HPA: tissue IHC). The mismatch raises concern for nonspecific binding or endogenous detection activity; it does not, by itself, identify which occurred. |
| Brown signal spreads across cells and surrounding tissue, obscuring cell boundaries. | The result cannot be scored reliably by cell type or compartment against the HPA profile (HPA: tissue IHC). General IHC practice: assess a primary-omission control and background outside cells before interpreting widespread chromogen as target staining. |
| A bronchus section shows no staining in basal cells. | Basal cells are a High-staining reference in HPA's tissue IHC data (HPA: High in bronchial basal cells). A negative run needs technical review before it supports a biological absence claim; HPA's Approved rating still has medium consistency with RNA data (HPA: tissue IHC). |
💡Expected UBE2I appearanceCall the result positive when the expected cells show interpretable nuclear and cytoplasmic staining, particularly High staining in bronchial basal or placental trophoblastic cells (HPA: tissue IHC); membrane-only signal is suspect (UniProt P63279: topology).
| Cell type within a tissue | Choose and score a defined cell population: HPA reports High staining in bronchial basal cells but Not detected in adipocytes, cholangiocytes and skeletal-muscle myocytes (HPA: tissue IHC). A whole-section average can hide that distinction. |
| Expected compartment | Tissue IHC is described as nuclear and cytoplasmic (HPA: tissue IHC); UniProt also lists a perinuclear region and describes UBE2I as mainly nuclear by similarity (UniProt P63279: subcellular location). Do not require exclusively nuclear staining. |
| IHC antibody evidence | HPA marks HPA003909 and CAB009021 Approved for IHC (HPA: antibodies). Its tissue profile has medium consistency with RNA data (HPA: tissue IHC). These ratings support an expected pattern but do not establish that every stained cell is specific. |
| Antigen retrieval and dilution | No UBE2I-specific retrieval condition or antibody dilution is supplied in the UniProt and HPA payloads. General IHC practice: use the selected antibody's documented IHC-P conditions as the starting point, then assess control sections. |
| Q: What should IF/ICC show? | A: Mainly nucleoplasm, with additional nucleolar localization in HPA's ICC-IF data; images are listed for MCF-7 and U2OS (HPA: subcellular). That cell-image result does not replace the tissue IHC pattern (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in bronchial basal cells or placental trophoblastic cells. | Either cell type is a High-staining reference (HPA: tissue IHC); the blank result may reflect a failed stain, though that observation cannot identify the failed step. | General IHC practice: check the positive control, antibody dilution, retrieval record and detection reagents for the run; interpret the test section only after the control performs. |
| Strong signal appears only at cell membranes. | This conflicts with nuclear and cytoplasmic locations and the absence of a transmembrane segment (HPA: tissue IHC; UniProt P63279: topology). | General IHC practice: compare matched control sections, inspect chromogen deposits and repeat with an IHC-validated antibody if the pattern persists. |
| Adipocytes or skeletal-muscle myocytes stain strongly. | HPA lists both cell types as Not detected (HPA: tissue IHC). Unexpected signal could arise from nonspecific binding or detection background; the pattern alone cannot distinguish them. | General IHC practice: compare a primary-omission control and review blocking and detection steps, while scoring adjacent cell types separately. |
| A diffuse brown haze makes nuclei hard to assess. | Background can obscure the nuclear and cytoplasmic distribution described for tissue IHC (HPA: tissue IHC); appearance alone does not establish its source. | General IHC practice: inspect the primary-omission control, washing, blocking and detection exposure; adjust the step implicated by the control before rescoring. |
| A tissue appears weaker than the selected High-staining reference. | UBE2I staining varies by cell type: HPA reports Medium in cerebellar granular-layer cells and Low in pancreatic endocrine cells (HPA: tissue IHC). | Record the tissue and cell population before judging the run. Compare it with its own HPA category and a same-run positive control (HPA: tissue IHC; general IHC practice). |
| A positive section shows cytoplasmic signal without clear nuclear enrichment. | HPA's tissue description includes both compartments, while UniProt's mainly nuclear note is qualified as by similarity (HPA: tissue IHC; UniProt P63279: subcellular location). | Score compartments separately and compare expected positive cells with background controls; reserve an artifact call for a pattern the controls or tissue profile cannot support. |
Experiment evidence
IHC & IF images showing UBE2I staining
A02295 · IHCIHC analysis of UBE2I UBC9 using anti-UBE2I UBC9 antibody (A02295). UBE2I UBC9 was detected in paraffin-embedded section of human appendicit
A02295 · IHCIHC analysis of UBE2I UBC9 using anti-UBE2I UBC9 antibody (A02295). UBE2I UBC9 was detected in paraffin-embedded section of human lung cance
A02295 · IFIF analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295). UBE2I/UBC9 was detected in immunocytochemical section of U20S cell. Enzym
A02295-1 · IHCIHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1). UBE2I/UBC9 was detected in paraffin-embedded section of human colon ca
A02295-1 · IHCIHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1). UBE2I/UBC9 was detected in paraffin-embedded section of human colon ca
A02295-1 · IFIF analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1). UBE2I/UBC9 was detected in immunocytochemical section of A431 cells. En
M02295 · IHCIHC analysis of UBE2I using anti-UBE2I antibody (M02295). UBE2I was detected in a paraffin-embedded section of mouse brain tissue. Heat medi
M02295 · IHCIHC analysis of UBE2I using anti-UBE2I antibody (M02295). UBE2I was detected in a paraffin-embedded section of rat brain tissue. Heat mediat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000103275-UBE2I/tissue/cerebral+cortex#imid_2229564
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000103275-UBE2I/tissue/colon#imid_2229490
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000103275-UBE2I/tissue/liver#imid_2229491
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000103275-UBE2I/tissue/kidney#imid_2229492
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody CAB079920; green = UBE2I, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody CAB079920; green = UBE2I, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBE2I IHC & IF
🧪Run placenta first and expect staining in trophoblastic cells (HPA: High in trophoblastic cells). Use adipose tissue adipocytes as a negative comparison (HPA: Not detected in adipocytes); no other cell population on the placenta slide is validated here as an internal negative, so assess unstained neighboring cells for background without assuming they lack UBE2I (HPA: placenta row specifies trophoblastic cells only).
Positive control tissue: Bronchus (Basal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UBE2I in MCF-7, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit isotype control for the rabbit primary, plus a UBE2I knockout specimen or a validated peptide-block control (caption: rabbit anti-UBE2I; standard IHC practice). Check placental background from endogenous peroxidase and biotin when using the caption’s biotin-based DAB detection (caption: biotinylated secondary, SABC/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A02295 paraffin-section caption does not state a fixative (caption: fixative unreported). The caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes, which documents a tested condition without establishing that retrieval is required (caption: citrate retrieval). ICC-IF images are available for MCF-7 and U2OS, but the supplied evidence does not establish whether IF or frozen sections are easier; placental endogenous biotin can complicate the caption’s SABC detection (HPA: ICC-IF image cell lines; caption: SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for UBE2I
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Non-ciliated cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBE2I IHC Tips
Troubleshoot UBE2I staining in paraffin sections by checking retrieval, compartment patterns, controls and cell-level scoring before interpreting chromogenic signal.
What retrieval should I try when UBE2I staining is weak?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A02295). The selected paraffin-section example used this condition before overnight incubation with 1 μg/ml antibody at 4°C (datasheet A02295). If signal remains weak, check that sections were fully deparaffinised and compare retrieval duration on adjacent sections while keeping detection conditions constant (standard IHC practice). Assess nuclear and cytoplasmic staining in glandular cells of appendix, where expression is reported at a medium level, alongside a no-primary control (HPA: Appendix glandular cells, Medium; standard IHC practice). Avoid treating stronger diffuse staining alone as improved retrieval; evaluate compartment definition and background together (standard IHC practice).
Could fixation explain inconsistent UBE2I staining between paraffin blocks?
Target-specific fixation sensitivity for UBE2I is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (datasheet A02295). Record each block’s fixative, fixation duration and processing history before comparing staining intensity across specimens (standard IHC practice). On adjacent sections, keep citrate retrieval at pH 6 for 20 minutes and the detection workflow constant while comparing blocks (datasheet A02295; standard IHC practice). Check tissue preservation and nuclear detail with the counterstain, because damaged morphology can make compartment scoring unreliable (standard IHC practice). If a block stains weakly, compare an intact reference section in the same run before attributing the difference to fixation (standard IHC practice).
How should I troubleshoot cytoplasmic or perinuclear UBE2I staining?
UBE2I is reported in nucleus, cytoplasm and the perinuclear region, with a mainly nuclear annotation (UniProt P63279: subcellular location). In tissue IHC, nuclear and cytoplasmic expression occurs in most tissues, so cytoplasmic signal alone is not automatically erroneous (HPA: tissue profile). Compare stained cells with adjacent morphology, a no-primary control and sections processed in the same run before assigning a compartment pattern (standard IHC practice). If perinuclear staining appears only at section edges or alongside widespread chromogen haze, review section drying, blocking and detection background (standard IHC practice). Report nuclear and cytoplasmic scores separately when their distributions differ, rather than collapsing them into one intensity value (standard IHC practice).
Could an isoform or modified epitope explain discordant staining?
The supplied record lists 0 isoforms and a UBC core spanning residues 4–157, but it does not identify this antibody’s epitope (UniProt P63279: isoforms and domain; datasheet A02295). Reported modifications include acetylation at residues 2 and 65 and CDK1-associated phosphorylation at 71 (UniProt P63279: modified residues). These annotations do not establish that any modification changes binding in paraffin sections (UniProt P63279: modified residues; datasheet A02295: epitope unspecified). When staining differs across blocks, first compare retrieval, tissue preservation and control staining under matched processing (standard IHC practice). If epitope dependence matters, obtain antibody epitope information or compare an independently validated antibody before proposing a modification-specific explanation (standard IHC practice).
How can I check UBE2I localisation by multiplex IF?
Use IF as a separate assay and validate its staining controls independently of the paraffin-section chromogenic workflow (standard IF practice). UBE2I has no transmembrane segment and is mainly nuclear, with cytoplasmic and perinuclear annotations; include permeabilisation suitable for access to these intracellular compartments (UniProt P63279: topology and subcellular location; standard IF practice). Multiplex with a marker identifying the expected cell type, then inspect UBE2I within those marked cells rather than relying on tissue-wide fluorescence (standard IF practice). Choose spectrally separated fluorophores after checking tissue autofluorescence and include single-label controls for channel bleed-through (standard IF practice). Supported nucleoplasmic and additional nucleolar localisation provides a compartment reference for IF assessment (HPA: subcellular summary).
What should I check when DAB obscures UBE2I-positive cells?
The selected paraffin-section example used 10% goat serum blocking, a biotinylated secondary antibody and DAB development (datasheet A02295). If background is diffuse, compare a no-primary section and inspect whether signal follows tissue structures or appears across empty spaces and section edges (standard IHC practice). Apply an appropriate peroxidase block before chromogenic detection and assess secondary or avidin-biotin background with matching controls (standard IHC practice). Titrate development and antibody exposure against a reference section while preserving visible nuclear detail in the counterstain (standard IHC practice). Judge improvement by clearer cell boundaries and expected compartment staining, not simply by a paler slide (standard IHC practice; UniProt P63279: subcellular location).
How should I score UBE2I staining across cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because UBE2I shows nuclear and cytoplasmic expression in most tissues (HPA: tissue profile). For nuclear or cytoplasmic compartments separately, record the percentage of positive cells and staining intensity, then calculate an H-score when its intensity categories are predefined (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable, annotated tissue and normalise counts to that measured area (standard IHC practice). Exclude folds, necrosis and poorly preserved regions using a rule set before reviewing group identities (standard IHC practice). Keep retrieval, DAB development and image thresholds consistent, and report compartments and cell types with the resulting scores (standard IHC practice).
How do I distinguish genuine UBE2I signal from artefact?
A plausible UBE2I pattern includes nuclear and cytoplasmic staining, while the protein is annotated as mainly nuclear (HPA: tissue profile; UniProt P63279: subcellular location). Compare the signal with cell identity: appendix glandular cells show medium staining, whereas adipocytes and skeletal-muscle myocytes are listed as not detected (HPA: Appendix, Adipose tissue and Skeletal muscle). Repeated signal at cut edges, within necrosis or without intact cell boundaries warrants an artefact check before scoring (standard IHC practice). Persistent DAB in a no-primary control warrants review of endogenous enzyme activity and the detection reagents (standard IHC practice). Interpret unexpected compartments or cell populations only after examining morphology, matched controls and independently processed sections (standard IHC practice).
Boster reagents
Best UBE2I / SUMO-conjugating enzyme UBC9 IHC Antibodies
The catalog antibodies have IHC images from human paraffin sections and mouse and rat brain sections, plus IF/ICC images from human cells (catalog image captions).
Real IHC data
Anti-UBE2I UBC9 Antibody ®
Cat # A02295
Real IHC data
Anti-UBE2I/UBC9 Antibody ®
Cat # A02295-1
Real IHC data
Anti-UBE2I/Ubc9 Rabbit Monoclonal Antibody
Cat # M02295
A02295 has IHC images from human appendicitis and lung cancer sections and mouse and rat brain sections, plus IF/ICC data from U20S cells (catalog image captions). A02295-1 has IHC images from human colon cancer, mammary cancer and tonsil sections and IF/ICC data from A431 cells; M02295 has IHC images from mouse and rat brain sections (catalog image captions).
Which to pick: For human tissue IHC, consider A02295-1, whose own images show paraffin sections of colon cancer, mammary cancer and tonsil; for mouse or rat brain sections, consider monoclonal M02295, whose own images show paraffin brain sections (catalog image captions; catalog: M02295 clone EDF-21). For IF/ICC, A02295-1 has an A431 cell image and A02295 has a U20S cell image (catalog IF image captions). For IHC across human, mouse and rat samples, A02295 has paraffin-section images in all three species; the captions do not report the fixative (catalog IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P63279 (UBC9_HUMAN, SUMO-conjugating enzyme UBC9).
- Human Protein Atlas. UBE2I tissue IHC expression (reliability: Approved).
- Human Protein Atlas. UBE2I subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli..
- Human Protein Atlas. UBE2I antibody validation summary (3 antibodies).
- Identification of UBE2I as a Novel Biomarker in ccRCC Based on a Large-Scale CRISPR-Cas9 Screening Database and Immunohistochemistry. Frontiers in molecular biosciences 2022 — PMC8861443.
- UBC9-Mediated SUMO Pathway Drives Prohibitin-1 Nuclear Accumulation and PITX1 Repression in Primary Osteoarthritis. International journal of molecular sciences 2025 — PMC12249805.
- Targeting UBC9-mediated protein hyper-SUMOylation in cystic cholangiocytes halts polycystic liver disease in experimental models. Journal of hepatology 2021 — PMC8157180.
- Increased PD-L1 Restricts Liver Injury in Nonalcoholic Fatty Liver Disease. Oxidative medicine and cellular longevity 2022 — PMC9126662.
- PubMed PMID:8668529 — UniProt-cited evidence.
- PubMed PMID:9067428 — UniProt-cited evidence.
- PubMed PMID:8565643 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBE2I (SUMO-conjugating enzyme UBC9). Expected localisation Nuclear and cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02295). Validated IHC antibody: A02295 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Basal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendicitis tissue; fixative not specified (datasheet A02295); 2. Fixation: Image fixative and duration unreported (datasheet A02295); 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 A02295); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02295); 8. Primary antibody: Rabbit anti-UBE2I, 0.5-1μg/ml (datasheet A02295); 9. Primary incubation: Overnight at 4 °C (datasheet A02295). Watch for: Fixation: Keep fixation consistent across paraffin sections and controls. (selected-SKU IHC image A02295); Caveat: Staining has only medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms listed; the mature chain spans residues 2–158 (UniProt); Regulation: No specific expression regulator is reported (UniProt). Evidence: UniProt P63279, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A02295, PMC8861443. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBE2I/immunohistochemistry-ihc-guide.