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UBE2G1 / Ubiquitin-conjugating enzyme E2 G1 · IHC design guide
Design Immunohistochemistry for UBE2G1
Plan chromogenic IHC for UBE2G1 in paraffin sections using the catalog antibody's 1:100–1:300 dilution range (datasheet A09806-1). Use the reported cytoplasmic pattern and high-staining glandular and hematopoietic cells to assess staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBE2G1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in glandular and hematopoietic cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09806-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Mainly expressed in skeletal muscle (UniProt) | |
| Isoform / epitope | No isoforms; Met1 removal warrants an N-terminal epitope check (UniProt) |
Section 1
Recommended UBE2G1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A09806-1) is accompanied by one published UBE2G1 IHC protocol in large yellow croaker (PMC9368838).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A09806-1) |
| Fixation | Image fixative and duration unreported (datasheet A09806-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A09806-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-UBE2G1, 1:100-1:300 (datasheet A09806-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBE2G1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 heat retrieval for the catalog antibody (datasheet A09806-1); the published protocol does not specify retrieval (PMC9368838).
Section 2
What Is the Expected UBE2G1 Staining Pattern?
Expect predominantly cytoplasmic UBE2G1 staining in paraffin sections (HPA tissue IHC: general cytoplasmic expression). High staining is reported in glandular cells of the adrenal gland, appendix, colon and duodenum, among other cell populations (HPA tissue IHC). UBE2G1 has no transmembrane segment (UniProt P62253 topology). HPA rates its tissue IHC profile Approved, with medium agreement between staining and RNA data; external verification is pending (HPA tissue IHC).
| Cytoplasmic chromogen in glandular cells of colon or duodenum, with discernible cell boundaries. | This fits the reported compartment and cell types; HPA scores these glandular cells High (HPA tissue IHC). Compare staining with the section’s morphology and controls. An HPA level describes an observed pattern, not a required intensity for every cell or staining run. |
| Signal confined to cell membranes, lumens or extracellular material, with little cytoplasmic staining. | That distribution conflicts with general cytoplasmic tissue staining (HPA tissue IHC) and the absence of a transmembrane segment (UniProt P62253 topology). Check section morphology and detection controls before treating the pattern as UBE2G1. |
| Strong chromogen in unexpected cells while an expected cell population is weak. | Investigate antibody cross-reactivity or endogenous detection activity. UBE2G1 is widely expressed (UniProt P62253 tissue specificity), so an unexpected cell type alone cannot prove a false positive. Compare cell-specific staining with HPA observations and appropriate controls. |
| Diffuse colour covers tissue, empty spaces and cell-free areas. | Poorly localized colour makes the cytoplasmic pattern difficult to assess (HPA tissue IHC: general cytoplasmic expression). As general IHC practice, inspect reagent-only or primary-omission controls, blocking and wash steps for background before scoring cells. |
| No staining in a colon glandular-cell section expected to be positive. | HPA reports High staining in colon glandular cells (HPA tissue IHC), making them a useful comparison. First verify that the relevant cells are present and the detection control worked; an absent signal alone does not establish absent UBE2G1. |
💡Expected UBE2G1 appearanceA positive IHC result shows predominantly cytoplasmic chromogen in the relevant cells, including glandular cells scored High in colon (HPA tissue IHC); isolated membrane, luminal or extracellular colour without that cellular pattern warrants investigation (HPA tissue IHC; UniProt P62253 topology).
| Tissue and cell selection | HPA reports High staining in adrenal, appendix, colon and duodenum glandular cells; bone marrow hematopoietic cells; caudate neuronal cells; and cerebellar granular-layer cells (HPA tissue IHC). Select a comparison section whose expected cells can be identified morphologically. |
| Skeletal muscle expectation | UniProt describes widespread expression, mainly in skeletal muscle (UniProt P62253 tissue specificity), and HPA calls skeletal-muscle RNA tissue-enhanced (HPA tissue IHC). The supplied HPA tissue IHC observations give no skeletal-muscle staining level, so they do not establish its expected chromogen intensity. |
| Topology and antigen interpretation | UBE2G1 has no transmembrane segment and has a UBC core at residues 5–166 (UniProt P62253 topology/domains). The supplied record gives no antibody epitope, so these facts cannot predict whether a particular retrieval condition will expose it. |
| Evidence strength | HPA lists HPA045681 and HPA050551 as IHC Approved (HPA antibodies). The tissue profile has medium agreement with RNA data and awaits external verification (HPA tissue IHC). Treat the reported distribution as a reference for interpretation, with controls for the assay at hand. |
| IF/ICC Q: Should nuclear fluorescence be treated as the expected IHC result? | A: HPA reports approved nucleoplasm and cytosol localization by ICC-IF (HPA subcellular). For paraffin IHC, use its reported general cytoplasmic pattern as the primary comparison (HPA tissue IHC); assess any nuclear chromogen with controls rather than automatically calling it artefact. |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon glandular cells lack signal. | The expected cells may be absent from the examined area, or an assay step may have failed; HPA reports High staining in these cells (HPA tissue IHC). | Locate intact glands, check the positive and detection controls, then review retrieval, primary-antibody dilution and incubation as general IHC troubleshooting. No UBE2G1-specific retrieval or dilution condition is supplied. |
| A strong membrane rim replaces cytoplasmic staining. | The pattern differs from general cytoplasmic tissue staining (HPA tissue IHC) and lacks support from UBE2G1 topology (UniProt P62253: no transmembrane segment). | Inspect morphology and controls; reassess antibody-specific staining and chromogen deposition before assigning the rim to UBE2G1. |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible, but widespread expression limits conclusions from cell identity alone (UniProt P62253 tissue specificity). | Compare the same section with primary-omission and detection controls, then check whether cytoplasmic staining also appears in the HPA-reported cell populations (HPA tissue IHC). |
| Colour is diffuse across tissue and blank areas. | Background from detection reagents or insufficient blocking or washing can obscure cellular localization; the reference tissue pattern is cytoplasmic (HPA tissue IHC). | Use primary-omission and reagent controls, inspect blank areas, and adjust blocking or washing according to the assay’s general IHC workflow before scoring intensity. |
| Nuclear chromogen appears alongside cytoplasmic staining. | Nucleoplasm and cytosol are both approved ICC-IF locations (HPA subcellular), while the supplied tissue IHC profile describes general cytoplasmic expression (HPA tissue IHC). | Record the compartments separately and review controls and morphology. Do not infer that ICC-IF localization alone validates a nuclear paraffin-IHC signal. |
| Signal differs between two antibodies or sections. | HPA lists both supplied antibodies as IHC Approved, while the tissue profile has medium staining–RNA agreement and awaits external verification (HPA antibodies; HPA tissue IHC). | Compare the same cell populations and controls across sections, document any compartment difference, and avoid treating one discrepant stain as definitive localization. |
Experiment evidence
IHC & IF images showing UBE2G1 staining
A09806-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132388-UBE2G1/tissue/cerebral+cortex#imid_14051114
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132388-UBE2G1/tissue/colon#imid_14046000
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132388-UBE2G1/tissue/liver#imid_14045993
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000132388-UBE2G1/tissue/kidney#imid_14046002
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA050551; green = UBE2G1, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA050551; green = UBE2G1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBE2G1 IHC & IF
🧪Run adrenal gland first: glandular cells should stain strongly (HPA: High in adrenal gland glandular cells). HPA detects UBE2G1 in all 44 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells with little or no signal on the positive slide as background references rather than validated internal negatives (HPA: no negative rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: UBE2G1 is detected in all 44 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 UBE2G1 in Hep-G2, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host species and clonality, and a UBE2G1 knockout control or peptide block if the immunizing peptide is available (standard IHC practice). In adrenal sections, check pigment and vascular background, and quench endogenous peroxidase for chromogenic detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A09806-1 paraffin-section caption leaves the fixative unreported (datasheet caption). The caption uses Tris-EDTA at pH 9.0 for retrieval and a 1:200 primary dilution overnight at 4°C, but does not establish that this retrieval is required (datasheet caption). For IF/ICC, nucleoplasmic and cytosolic staining is a localization reference (HPA subcellular); whether IF/ICC or frozen sections are easier is unreported.
HPA tissue IHC evidence for UBE2G1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: UBE2G1 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced UBE2G1 IHC Tips
Troubleshoot UBE2G1 staining in paraffin sections by checking retrieval, staining controls, compartment patterns and cell-specific scoring.
What retrieval should I use when UBE2G1 staining is weak in paraffin sections?
Use heat-mediated Tris-EDTA at pH 9.0 for paraffin sections (datasheet A09806-1). The catalog antibody's human tonsil image used this retrieval with primary antibody at 1:200, incubated at 4°C overnight (caption A09806-1). If staining is weak, compare a consistently heated section with a matched section that received a shorter heating period; excessive heating can damage morphology (standard IHC practice). Keep cooling and wash conditions consistent while assessing whether glandular or hematopoietic cells develop interpretable cytoplasmic staining (HPA: high in colon glandular cells and bone marrow hematopoietic cells; HPA: general cytoplasmic expression). Record the fixative separately because the caption identifies paraffin embedding but does not state fixation (caption A09806-1).
Could fixation explain weak or uneven UBE2G1 IHC staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not identify a fixative (caption A09806-1). Record each specimen's fixative and fixation duration, then compare sections processed with the same retrieval, antibody incubation and detection conditions; fixation can alter antigen accessibility in IHC (standard IHC practice). Use the documented pH 9.0 Tris-EDTA retrieval and 1:200 primary dilution as reference conditions when assessing the catalog antibody (caption A09806-1). Examine morphology alongside signal, since poor preservation can make cellular assignment unreliable (standard IHC practice). Do not infer a UBE2G1-specific fixation effect from its tissue distribution or sequence features (HPA: tissue IHC profile; UniProt P62253: protein record).
Should UBE2G1 staining appear in cytoplasm, nucleus or at membranes?
Expect a predominantly cytoplasmic pattern in tissue IHC (HPA: general cytoplasmic expression). Nucleoplasmic and cytosolic localisation are both approved in cell imaging, so a nuclear component warrants assessment rather than automatic rejection (HPA: approved nucleoplasm and cytosol). UniProt does not annotate a subcellular location, while the protein has no transmembrane segment; a crisp membrane-only pattern therefore needs independent validation (UniProt P62253: subcellular annotation and topology). Compare the same cell populations across sections and include a no-primary control to assess nonspecific detection (standard IHC practice). Judge weak nuclear signal against counterstain and morphology before assigning it to UBE2G1 (standard IHC practice).
How can UBE2G1 sequence features affect epitope interpretation in IHC?
The record lists 0 isoforms and a UBC core spanning residues 5–166 (UniProt P62253: isoforms and domain). It also lists full-length residues 1–170 and an N-terminally processed chain spanning 2–170, with alternative N-terminal acetylation annotations (UniProt P62253: processing and modified residues). If the antibody's mapped epitope is available, check whether it overlaps that N-terminal region before interpreting a staining difference as altered abundance (standard IHC interpretation; UniProt P62253: processing). An unmapped epitope does not establish which processed form is detected (standard antibody interpretation). Compare matched sections using the documented pH 9.0 retrieval before attributing a changed pattern to epitope accessibility (caption A09806-1; standard IHC practice).
How should IF help investigate an ambiguous UBE2G1 IHC pattern?
Use IF as a localisation check alongside the chromogenic IHC result: cell imaging reports approved nucleoplasmic and cytosolic UBE2G1 signal (HPA: subcellular profile). Multiplex with a validated marker for the cell population being examined, such as glandular cells in colon, and include single-channel controls (HPA: high in colon glandular cells; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence and separating their spectra from the marker channel (standard IF practice). Because both expected compartments are inside the cell, assess permeabilisation sufficient for cytosolic and nucleoplasmic access while preserving morphology (HPA: approved locations; standard IF practice). The 1:200 dilution belongs to the selected IHC image and is not an IF setting (caption A09806-1).
How can I distinguish UBE2G1 signal from chromogenic background?
Run a no-primary control and inspect unstained tissue pigment before interpreting diffuse brown deposits as UBE2G1 (standard IHC practice). For a peroxidase and DAB workflow, block endogenous peroxidase and check whether residual colour persists in the control; these are general detection steps (standard IHC practice). The selected tonsil image used primary and secondary antibodies at 1:200, with the secondary incubated for 30 minutes at room temperature (caption A09806-1). If background rises across all compartments, review blocking, washes and antibody concentration on matched sections (standard IHC practice). Compare cellular signal with the reported general cytoplasmic tissue pattern, while allowing for a nuclear component supported by cell imaging (HPA: tissue IHC and subcellular profiles).
What is a reproducible way to score UBE2G1 chromogenic IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC reports general cytoplasmic expression while cell imaging also supports nucleoplasmic localisation (HPA: tissue IHC and subcellular profiles). For intact cells, report percent positive and an H-score using intensity categories 0–3, yielding a 0–300 scale (standard IHC scoring practice). If spatial distribution is the question, report positive-cell density per mm² of viable tissue and state how cells and area were counted (standard IHC quantification practice). Normalize comparisons to the same eligible cell type or viable tissue area, and exclude folds and necrosis (standard IHC practice). Score matched processing batches with the same threshold and retain no-primary controls for background assessment (standard IHC practice).
Which UBE2G1 staining patterns merit validation before calling them positive?
Prioritize cellular staining consistent with the general cytoplasmic tissue profile, while allowing a nucleoplasmic component supported by cell imaging (HPA: tissue IHC and approved subcellular locations). A membrane-only outline deserves validation because UBE2G1 lacks a transmembrane segment (UniProt P62253: topology). Check whether staining follows the expected cell population: high signal is reported in colon glandular cells and bone marrow hematopoietic cells (HPA: tissue IHC). Disregard edge-heavy deposits, necrotic areas and colour retained in a no-primary control; endogenous peroxidase can confound DAB interpretation (standard IHC practice). Treat agreement with HPA as supportive rather than conclusive because its tissue profile is approved with medium staining-to-RNA consistency and awaits external verification (HPA: reliability).
Boster reagents
Best UBE2G1 / Ubiquitin-conjugating enzyme E2 G1 IHC Antibodies
A09806-1 has a human tonsil paraffin-section IHC image (IHC image caption) and lists IF and human, mouse and rat reactivity (catalog: applications; catalog: reactivity).
Real IHC data
Anti-UBE2G1 Antibody
Cat # A09806-1
A09806-1 was imaged by IHC on paraffin-embedded human tonsil at 1:200, following Tris-EDTA retrieval at pH 9.0 (IHC image caption). It lists IF and human, mouse and rat reactivity, but supplies no IF image (catalog: applications; catalog: reactivity; catalog: IF image alts).
Which to pick: For tissue IHC, choose A09806-1 for paraffin sections using the human tonsil example as a starting point; the fixative is unreported (IHC image caption). For IF/ICC, A09806-1 lists IF at 1:50, but ICC is not listed and no IF image is supplied; its host is rabbit and clonality is unreported (catalog: applications; catalog: IF dilution; catalog: IF image alts; catalog: host and clone). For cross-species work, A09806-1 lists human, mouse and rat reactivity, while its supplied IHC image documents human tissue only (catalog: reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62253 (UB2G1_HUMAN, Ubiquitin-conjugating enzyme E2 G1).
- Human Protein Atlas. UBE2G1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. UBE2G1 subcellular location (ICC-IF): Localized to the nucleoplasm and cytosol..
- Human Protein Atlas. UBE2G1 antibody validation summary (2 antibodies).
- UBE2G1 Is a Critical Component of Immune Response to the Infection of Pseudomonas Plecoglossicida in Large Yellow Croaker (Larimichthys crocea). International journal of molecular sciences 2022 — PMC9368838.
- PubMed PMID:8893823 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBE2G1 (Ubiquitin-conjugating enzyme E2 G1). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A09806-1). Validated IHC antibody: A09806-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 44 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A09806-1); 2. Fixation: Image fixative and duration unreported (datasheet A09806-1); 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: Tris-EDTA pH 9.0 (datasheet A09806-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-UBE2G1, 1:100-1:300 (datasheet A09806-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms; Met1 removal warrants an N-terminal epitope check (UniProt); Regulation: Mainly expressed in skeletal muscle (UniProt). Evidence: UniProt P62253, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and cytosol., Datasheet A09806-1, PMC9368838. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBE2G1/immunohistochemistry-ihc-guide.