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UBA1 / Ubiquitin-like modifier-activating enzyme 1 · IHC design guide
Design Immunohistochemistry for UBA1
Plan UBA1 paraffin-section IHC around the nuclear staining seen in most tissues (HPA tissue IHC). This guide helps interpret high staining in glandular and kidney tubular cells alongside low staining in neuronal and muscle cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBA1 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M02810) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M02810) | |
| Caveat | Low neuronal and muscle staining may be missed (HPA tissue IHC) | |
| Regulation | No specific expression regulation reported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage unreported (UniProt) |
Section 1
Recommended UBA1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses heat-mediated EDTA retrieval at pH 8.0 (datasheet M02810). One published paraffin-section IHC protocol provides a citrate-retrieval alternative (PMC7744729).
| Sample | Paraffin-embedded human acinic cell carcinoma of parotid tissue; fixative not specified (datasheet M02810) |
| Fixation | Image fixative and duration unreported (datasheet M02810); 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 M02810); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M02810) |
| Primary antibody | Rabbit monoclonal (clone IAC-21) anti-UBA1, 1:50 (datasheet M02810) |
| Primary incubation | Overnight at 4 °C (datasheet M02810) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M02810) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | UBA1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet M02810); consider the published citrate condition when adapting the HCC protocol (PMC7744729).
Section 2
What Is the Expected UBA1 Staining Pattern?
UBA1 should show predominantly nuclear staining across most tissues (HPA tissue IHC: Approved; medium consistency with RNA). High staining is reported in adrenal glandular cells, bronchial respiratory epithelial cells, cerebellar Purkinje cells, and kidney tubule cells (HPA tissue IHC: High). Nucleoplasmic localisation with additional cytosolic signal is supported by ICC-IF (HPA subcellular). UBA1 also has cytoplasmic, mitochondrial, and nuclear annotations and no transmembrane segment (UniProt P22314).
| Clear nuclear chromogen in kidney tubule cells or bronchial respiratory epithelial cells. | This matches high staining in those cells and the predominant nuclear tissue pattern (HPA tissue IHC: High; nuclear in most tissues). Assess cell identity and nuclear localisation together. |
| Predominantly membranous, extracellular, or vessel-lumen staining with little nuclear signal. | Treat this as suspect: UBA1 has no transmembrane segment, and HPA reports a mainly nuclear pattern (UniProt P22314 topology; HPA tissue IHC). Check whether staining follows tissue edges or deposits. |
| Strong staining in a cell population outside the reported high-staining cells. | Cross-reactivity or endogenous chromogenic activity may contribute (general IHC practice). HPA lists high-staining populations but supplies no negative cell population; unexpected staining alone does not establish absence of UBA1 (HPA tissue IHC). |
| Uniform haze across nuclei, cytoplasm, and tissue-free areas. | Diffuse background obscures compartment assignment (general IHC practice). Compare the signal with a detection-only control before scoring UBA1 localisation; the expected tissue pattern is mainly nuclear (HPA tissue IHC). |
| No nuclear signal in a reported high-staining cell population. | An uninformative stain is possible before biological absence is inferred (general IHC practice). Confirm that the named cells are present and inspect controls; HPA reports high staining in kidney tubules, among other populations (HPA tissue IHC: High). |
💡Expected UBA1 appearanceCall a result positive when identifiable, reported high-staining cells show clear predominantly nuclear chromogen (HPA tissue IHC: High; nuclear in most tissues); diffuse haze or chiefly membranous staining is suspect (general IHC practice; UniProt P22314 topology).
| Compartment assignment | Tissue IHC is predominantly nuclear; ICC-IF supports nucleoplasm plus cytosol (HPA tissue IHC; HPA subcellular). Some cytoplasmic signal is plausible, but chromogenic IHC alone cannot resolve a mitochondrial contribution (UniProt P22314 localisation; general IHC practice). |
| Tissue and cell selection | HPA reports high staining in specified glandular, epithelial, Purkinje, and tubule cells, but low staining in hippocampal neurons, cardiomyocytes, and skeletal myocytes (HPA tissue IHC). Use the named cell population when judging intensity. |
| Strength of tissue evidence | The tissue profile is Approved with medium consistency between staining and RNA (HPA tissue IHC). Three listed antibodies have Approved IHC status, but that status alone does not make every unusual field UBA1-specific (HPA antibodies; general IHC interpretation). |
| Protein forms and topology | UniProt lists two isoforms, no signal peptide or propeptide, and no transmembrane segment (UniProt P22314). These annotations do not identify which form an unspecified antibody detects or justify a membrane staining call. |
| Chromogenic detection background | Endogenous enzyme activity can mimic deposited chromogen, depending on the detection system (general IHC practice). A detection-only control helps separate that activity from antibody-dependent signal; HPA supplies no UBA1-specific background rate. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining reference cells lack nuclear signal. | The run or tissue may be uninformative; absence is difficult to interpret where HPA reports high staining (HPA tissue IHC: High; general IHC practice). | Verify that the named cells are present, then review section integrity, antigen retrieval, antibody incubation, and detection controls (general IHC practice). Change one workflow variable at a time. |
| Chromogen outlines membranes or lies outside cells. | This conflicts with the mainly nuclear tissue pattern and lack of a transmembrane segment (HPA tissue IHC; UniProt P22314 topology). | Inspect tissue morphology and a detection-only control; score UBA1 only where cellular staining has a credible compartment pattern (general IHC practice). |
| Every compartment has brown haze. | Nonspecific antibody binding or chromogenic background can obscure nuclear localisation (general IHC practice). | Compare detection-only and antibody-stained sections; review blocking, antibody concentration, washing, and counterstain before interpreting intensity (general IHC practice). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). HPA provides low-staining examples but no negative population (HPA tissue IHC). | Check the cell identity and control staining; compare with a reported high-staining population, and avoid calling the unexpected cells definitively positive from one field (HPA tissue IHC; general IHC practice). |
| Heart or skeletal muscle appears faint beside a strong reference. | Low staining is reported in cardiomyocytes and skeletal myocytes (HPA tissue IHC: Low); faint signal alone does not show a failed run. | Judge run performance in a reported high-staining population and score the low-staining population separately (HPA tissue IHC; general IHC practice). |
| Does IF/ICC require the same compartment call as tissue IHC? | The assays provide different resolution: tissue IHC is mainly nuclear, while ICC-IF supports nucleoplasm plus cytosol (HPA tissue IHC; HPA subcellular). | For IF/ICC interpretation, allow supported cytosolic signal alongside nucleoplasmic signal (HPA subcellular). Use the separate IF/ICC guide for its workflow. |
Experiment evidence
IHC & IF images showing UBA1 staining
M02810 · IHCIHC analysis of UBA1 using anti-UBA1 antibody (M02810). UBA1 was detected in a paraffin-embedded section of human acinic cell carcinoma of p
M02810 · IHCIHC analysis of UBA1 using anti-UBA1 antibody (M02810). UBA1 was detected in a paraffin-embedded section of human acinic cell carcinoma of p
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130985-UBA1/tissue/cerebral+cortex#imid_5076596
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130985-UBA1/tissue/colon#imid_5076514
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130985-UBA1/tissue/liver#imid_5076515
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130985-UBA1/tissue/kidney#imid_5076516
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001506; green = UBA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001506; green = UBA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBA1 IHC & IF
🧪Run adrenal gland first; glandular cells should stain for UBA1 (HPA: High in adrenal gland glandular cells). HPA detects UBA1 in all 44 scored tissues, so there is no supported negative tissue or known UBA1-negative cell population on this slide; use no-primary and isotype controls to establish the background appearance (HPA: no negative rows; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: UBA1 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 UBA1 in A-431, U-251MG, U2OS, SiHa, RPTEC/TERT1, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only section and a concentration-matched rabbit isotype section alongside the primary-antibody section (caption: rabbit anti-UBA1; standard IHC practice). UBA1 knockout material can provide a biological specificity control if available; quench endogenous peroxidase and inspect unstained tissue for pigment that could be mistaken for DAB signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected M02810 paraffin-section caption does not state the fixative (M02810 caption: fixative unreported). The caption demonstrates staining after heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required (M02810 caption). There is no supplied comparison showing that frozen sections or IF are easier; HPA ICC-IF images support nucleoplasmic and cytosolic localisation, while endogenous peroxidase or pigment can complicate chromogenic interpretation (HPA: subcellular ICC-IF; standard IHC practice).
HPA tissue IHC evidence for UBA1
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: UBA1 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced UBA1 IHC Tips
Troubleshoot UBA1 staining in paraffin sections by checking retrieval, compartment patterns and cell level controls before interpreting chromogenic signal.
What retrieval should I try when UBA1 staining is weak in paraffin sections?
Start with heat mediated antigen retrieval in EDTA at pH 8.0 (datasheet M02810). The selected paraffin section image used this retrieval before incubation with M02810 at 1:50 overnight at 4°C (datasheet M02810). If signal remains weak, check deparaffinization, heating consistency and primary antibody coverage before adjusting retrieval duration on matched sections (standard IHC practice). Compare each adjusted section with a known positive section processed in the same run, since staining differences can arise from handling (standard IHC practice). Score nuclear and cytoplasmic staining separately: UBA1 has documented nuclear and cytoplasmic locations (UniProt P22314; HPA subcellular).
Could fixation explain inconsistent UBA1 staining across paraffin blocks?
The selected M02810 caption identifies a paraffin section but does not state its fixative, so UBA1 sensitivity to fixation is unknown here (datasheet M02810). Record each block’s fixative and processing history, then compare similarly handled sections within the same staining run (standard IHC practice). Keep retrieval at EDTA pH 8.0 while investigating fixation as a variable, because that is the documented M02810 retrieval condition (datasheet M02810). Check whether differences persist in morphologically intact regions and with consistent antibody incubation and detection (standard IHC practice). Do not assign a fixation effect from tissue staining patterns or UBA1’s annotated locations and modifications (HPA tissue IHC; UniProt P22314).
How should I assess nuclear versus cytoplasmic UBA1 staining?
Expect a substantial nuclear component: tissue IHC reports nuclear expression in most tissues, while subcellular imaging supports nucleoplasm and cytosol (HPA tissue IHC; HPA subcellular). UniProt also annotates cytoplasm, nucleus and mitochondrion for UBA1 (UniProt P22314). Score nuclear and cytoplasmic DAB separately in intact cells, using morphology and a counterstain to locate nuclei (standard IHC practice). If one compartment disappears after changing retrieval or detection conditions, compare matched sections before interpreting the shift biologically (standard IHC practice). A purely membranous rim warrants investigation, because UBA1 has no annotated transmembrane segment (UniProt P22314 topology).
Can this stain distinguish UBA1 isoforms or modification dependent epitopes?
UBA1 has 2 listed isoforms and several modified residues, including phosphorylated serines near the amino terminus (UniProt P22314). The supplied M02810 caption documents tissue staining conditions but gives no epitope map or isoform selectivity (datasheet M02810). Treat chromogenic signal as UBA1 immunoreactivity rather than assigning it to a particular isoform or modification without antibody specific validation (standard IHC practice). If cases differ unexpectedly, compare staining after the same EDTA pH 8.0 retrieval and 1:50 primary dilution (datasheet M02810). An altered staining pattern alone cannot establish which UBA1 epitope or modified residue was detected (standard IHC practice).
How can I examine UBA1 with a cell identity marker by IF?
For a separate IF experiment, pair UBA1 with a marker identifying the cell population being examined and assign clearly separated fluorophores (standard IF practice). Choose fluorophores after checking the section’s autofluorescence in each channel, and include single label controls when assessing overlap (standard IF practice). Use permeabilisation sufficient to expose intracellular epitopes: UBA1 is annotated in nucleus and cytoplasm and has no transmembrane segment (UniProt P22314 topology and subcellular location). Examine nucleoplasmic and cytosolic signal independently, consistent with supported subcellular imaging (HPA subcellular). The M02810 paraffin section caption documents chromogenic IHC conditions, so establish IF antibody performance with its own controls (datasheet M02810; standard IF practice).
What should I check when UBA1 DAB staining appears diffuse or uneven?
First compare the stained section with a no primary control to assess nonspecific detection and endogenous peroxidase signal (standard IHC practice). Check blocking and washing consistency; the M02810 tissue caption used 10% goat serum and a peroxidase conjugated secondary (datasheet M02810). Inspect tissue edges, folds and damaged regions separately, because uneven reagent access or trapped detection reagent can distort DAB staining (standard IHC practice). Recheck primary incubation against the documented 1:50 overnight at 4°C condition before changing antibody concentration (datasheet M02810). Diffuse cytoplasmic signal alone should not be rejected: cytosolic UBA1 is supported, alongside nucleoplasmic staining (HPA subcellular).
How should I quantify UBA1 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define a consistent cell population and compartment before scoring, since UBA1 has documented nuclear and cytoplasmic locations (UniProt P22314; HPA subcellular). Record the percentage of positive cells and staining intensity, or calculate a compartment specific H-score using the same thresholds across specimens (standard IHC practice). If counting positive cells per mm², normalize to evaluable tissue area and report cell density separately from the positive fraction (standard IHC practice). Exclude folds, necrosis and section edges using the same rules for every specimen (standard IHC practice). Keep imaging exposure, counterstain assessment and DAB scoring criteria consistent across runs (standard IHC practice).
How can I distinguish convincing UBA1 positivity from staining artefact?
Convincing UBA1 staining should be assessed in intact cells with attention to nuclei and cytoplasm, matching documented tissue and subcellular patterns (HPA tissue IHC; HPA subcellular). Compare candidate positive cells with their morphology and a relevant positive control; high staining is reported in kidney tubule cells and several glandular cell populations (HPA tissue IHC; standard IHC practice). Suspect artefact when signal concentrates at section edges, folds or necrotic areas, especially if a no primary control also stains (standard IHC practice). Check endogenous peroxidase contribution before interpreting DAB deposits as UBA1 (standard IHC practice). A predominantly membranous pattern needs further validation because UBA1 lacks an annotated transmembrane segment (UniProt P22314 topology).
Boster reagents
Best UBA1 / Ubiquitin-like modifier-activating enzyme 1 IHC Antibodies
The IHC-validated antibody has a paraffin-section image from human parotid acinic cell carcinoma (M02810 image caption). Human, mouse and rat reactivity and IF/ICC use are listed; no IF image is supplied (M02810 catalog).
Real IHC data
Anti-UBA1/Ube1 Rabbit Monoclonal Antibody
Cat # M02810
M02810 is the SKU with a rendered card and its own IHC figure, showing a human parotid acinic cell carcinoma paraffin section with EDTA retrieval and 1:50 primary antibody (M02810 image caption). Its catalog also lists IF/ICC and human, mouse and rat reactivity, but provides no IF image (M02810 catalog).
Which to pick: Choose M02810 for paraffin-section IHC because its own caption documents that preparation; the fixative is unreported (M02810 image caption). For IF/ICC or mouse and rat samples, both M02810 and M02810-2 are rabbit monoclonals with those applications and species listed, but neither has an IF image and M02810-2 has no IHC image in the supplied catalog (M02810 and M02810-2 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P22314 (UBA1_HUMAN, Ubiquitin-like modifier-activating enzyme 1).
- Human Protein Atlas. UBA1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. UBA1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. UBA1 antibody validation summary (4 antibodies).
- Ubiquitin-Activating Enzyme E1 (UBA1) as a Prognostic Biomarker and Therapeutic Target in Breast Cancer: Insights into Immune Infiltration and Functional Implications. International journal of molecular sciences 2024 — PMC11641640.
- Systemic restoration of UBA1 ameliorates disease in spinal muscular atrophy. JCI insight 2016 — PMC5033939.
- Ubiquitin-Like Modifier Activating Enzyme 1 as a Novel Diagnostic and Prognostic Indicator That Correlates With Ferroptosis and the Malignant Phenotypes of Liver Cancer Cells. Frontiers in oncology 2020 — PMC7744729.
- Inhibition of the Ubiquitin-Activating Enzyme UBA1 Suppresses Diet-Induced Atherosclerosis in Apolipoprotein E-Knockout Mice. Journal of immunology research 2020 — PMC7109586.
- PubMed PMID:1606621 — UniProt-cited evidence.
- PubMed PMID:1986373 — UniProt-cited evidence.
- PubMed PMID:1871145 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBA1 (Ubiquitin-like modifier-activating enzyme 1). Expected localisation Nuclear in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M02810). Validated IHC antibody: M02810 at 1:50 (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-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human acinic cell carcinoma of parotid tissue; fixative not specified (datasheet M02810); 2. Fixation: Image fixative and duration unreported (datasheet M02810); 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 M02810); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M02810); 8. Primary antibody: Rabbit monoclonal (clone IAC-21) anti-UBA1, 1:50 (datasheet M02810); 9. Primary incubation: Overnight at 4 °C (datasheet M02810). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M02810); Caveat: Low neuronal and muscle staining may be missed (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage unreported (UniProt); Regulation: No specific expression regulation reported (UniProt). Evidence: UniProt P22314, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M02810, PMC7744729. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBA1/immunohistochemistry-ihc-guide.