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UBA1 / Ubiquitin-like modifier-activating enzyme 1 · IHC design guide
Design Immunohistochemistry for UBA1
Plan chromogenic UBA1 IHC on paraffin sections around the nuclear pattern observed in most tissues (HPA tissue IHC). This guide covers fixation, antibody dilution, tissue controls and interpretation of low staining in hippocampal neurons and myocytes (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); cytoplasm and mitochondrion annotated (UniProt) | |
| 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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Hippocampal neurons and myocytes show low staining (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA RNA) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended UBA1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M02810) is accompanied by one published HCC IHC protocol (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 pH 8.0 retrieval for the catalog antibody (datasheet M02810); the HCC paper used sodium citrate (PMC7744729).
Section 2
What Is the Expected UBA1 Staining Pattern?
UBA1 is expected mainly in nuclei across most tissues (HPA tissue IHC: nuclear expression in most tissues; Approved, with medium consistency between staining and RNA data). Strong staining is reported in bronchial respiratory epithelium, kidney tubule cells and several glandular cell populations (HPA tissue IHC: High). UniProt also places UBA1 in the cytoplasm and mitochondrion and reports no transmembrane segment (UniProt P22314: subcellular location and topology).
| Predominantly nuclear staining in bronchial respiratory epithelial cells or kidney tubule cells. | This fits the reported nuclear tissue profile and High staining in these cells (HPA tissue IHC). Score the identified cell population, rather than treating every cell on the section as equally positive (general IHC practice). |
| Cytoplasmic signal accompanies a clear nuclear pattern. | Some cytoplasmic signal is plausible (UniProt P22314: cytoplasm; HPA ICC-IF: supported cytosol). An exclusively cytoplasmic IHC pattern departs from HPA's predominant nuclear tissue profile; review the staining and controls before interpreting it as UBA1 (HPA tissue IHC; general IHC practice). |
| Strong membrane or extracellular staining dominates the section. | This does not fit the reported intracellular locations or lack of a transmembrane segment (UniProt P22314: subcellular location and topology; HPA tissue IHC: nuclear profile). Treat it as a possible artefact or nonspecific signal and check controls (general IHC practice). |
| Unexpected cells stain strongly while the expected cell population is weak. | Compare cell identity and compartment with the HPA tissue profile; staining intensity varies by cell population (HPA tissue IHC). Cross-reactivity or endogenous chromogenic activity are possible explanations, not diagnoses from appearance alone (general IHC practice). |
| The whole section looks diffuse, or a reported High population has no signal. | Diffuse color can obscure cellular localization; absent signal in a reported high-staining population calls for a technical check (HPA tissue IHC; general IHC practice). Neither result alone establishes UBA1 absence (general IHC practice). |
💡Expected UBA1 appearanceCall a convincing positive when identifiable cells show predominantly nuclear staining, including strong signal in a reported High population such as bronchial respiratory epithelium or kidney tubules (HPA tissue IHC); dominant membrane staining or diffuse color without identifiable positive cells is suspect (UniProt P22314: topology; general IHC practice).
| Cell population and tissue | HPA reports High staining in selected epithelial, glandular, Purkinje and kidney tubule cells, but low staining in hippocampal neurons, cardiomyocytes and skeletal myocytes (HPA tissue IHC). Low is not a validated negative control. |
| Compartment used for scoring | Predominantly nuclear staining anchors tissue IHC interpretation (HPA tissue IHC). Cytoplasm and mitochondrion are also listed for UBA1 (UniProt P22314); the supplied tissue profile does not establish a separate mitochondrial IHC pattern. |
| Protein topology | UBA1 has no annotated transmembrane segment or signal peptide (UniProt P22314: topology and processing). A dominant cell-surface or extracellular pattern therefore needs scrutiny; this annotation does not identify the cause of such staining. |
| Antibody evidence | HPA marks HPA000289, CAB019435 and CAB073410 IHC Approved (HPA antibody validation). Its tissue profile has medium staining-to-RNA consistency (HPA tissue IHC); those labels do not prove every positive cell is specific. |
| Chromogenic detection | Endogenous enzyme activity can contribute color when the detection chemistry permits it (general IHC practice). Judge unexpected staining against assay controls and cell localization; HPA's UBA1 staining levels do not establish its cause. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a reported High cell population. | A detection or section-processing failure is possible (general IHC practice); the HPA level describes observed staining, not guaranteed performance in every run (HPA tissue IHC). | Check that the expected cells are present, then review the positive control, reagent sequence, retrieval conditions and detection system (general IHC practice). |
| Only cytoplasmic color is visible. | Cytoplasmic UBA1 is plausible (UniProt P22314), but this misses the predominant nuclear tissue pattern (HPA tissue IHC). Background or a compartment-scoring error is also possible (general IHC practice). | Compare nuclei with a counterstain and inspect a reported high-staining population; review controls before assigning the signal to UBA1 (HPA tissue IHC; general IHC practice). |
| Membrane or extracellular color dominates. | The pattern conflicts with UBA1's listed intracellular locations and lack of a transmembrane segment (UniProt P22314). | Check the negative control, wash performance and tissue morphology, then reassess whether any compartment-specific cellular signal remains (general IHC practice). |
| Unexpected cell types stain strongly. | Cross-reactivity or endogenous chromogenic activity may be involved (general IHC practice). HPA reports cell-specific variation and medium staining-to-RNA consistency (HPA tissue IHC). | Confirm cell identity on the counterstained section and compare the unexpected signal with assay controls and a reported high-staining population (HPA tissue IHC; general IHC practice). |
| Diffuse color makes nuclei hard to distinguish. | Background from detection chemistry or insufficient washing is possible (general IHC practice); diffuse color alone cannot establish UBA1 localization. | Inspect the negative control and review blocking, washing and detection conditions before scoring nuclei (general IHC practice). |
| Does IF/ICC require the same interpretation as tissue IHC? | HPA ICC-IF supports nucleoplasm as the main location and cytosol as an additional location (HPA subcellular). This is cellular localization evidence, separate from the tissue IHC intensity profile. | For IF/ICC, assess nucleoplasmic and cytosolic signals against the relevant channel controls (HPA subcellular; general IF practice). Use the separate IF/ICC guide for its assay design. |
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
🧪Start with adrenal gland: glandular cells should stain (HPA: High in adrenal gland glandular cells). HPA detects UBA1 in all 44 scored tissues, so there is no supported negative tissue or reliably negative cell type within the positive slide; use no-primary and isotype controls to define background, including in other cells on that slide (HPA: no negative tissue rows).
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 control and a concentration-matched rabbit isotype control alongside the slide (caption: rabbit primary antibody). A UBA1 knockout specimen can test target specificity; quench endogenous peroxidase and check for residual signal before interpreting HRP/DAB staining (caption: HRP/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected M02810 paraffin-section caption does not state the fixative (caption: fixative unreported). That caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or identify a tissue-specific artefact (caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF are easier; HPA documents ICC-IF localisation mainly in the nucleoplasm and also in the cytosol (HPA: subcellular ICC-IF).
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 chromogenic IHC by checking retrieval, tissue processing, compartmental staining, and controls before comparing signal across sections.
Which retrieval condition should I try first for UBA1 in paraffin sections?
Begin with heat-mediated EDTA retrieval at pH 8.0 for UBA1 paraffin-section IHC (datasheet M02810). The selected tissue image used this condition before 10% goat-serum blocking and 1:50 primary antibody overnight at 4°C (caption M02810). If staining is weak, adjust heating time on adjacent sections while keeping detection and antibody incubation constant (standard IHC practice). Compare nuclear signal in the test sections with a positive tissue control, since nuclear expression is reported in most tissues (HPA tissue IHC). Record tissue preservation and retrieval time with each run so a stronger signal is not mistaken for a biological difference (standard IHC practice).
How should I investigate weak UBA1 staining when tissue fixation varies?
The selected paraffin-section caption does not state a fixative, so UBA1-specific fixation sensitivity is unknown (caption M02810). Record fixation type and duration, then compare sections processed together with a documented positive control (standard IHC practice). Keep EDTA retrieval at pH 8.0 while assessing fixation, so retrieval changes do not confound the comparison (datasheet M02810; standard IHC practice). Score nuclear and cytoplasmic staining separately because UBA1 has both reported locations (UniProt P22314 subcellular; HPA subcellular). Treat any apparent loss after longer fixation as an observation requiring matched specimens and detection controls, rather than an established UBA1-specific effect (standard IHC practice).
Should UBA1 staining be nuclear, cytoplasmic, or both?
Expect predominantly nuclear staining across many tissues, while allowing cytoplasmic signal when controls support it (HPA tissue IHC; HPA subcellular). UBA1 is reported in the nucleus and cytoplasm, and cell imaging supports nucleoplasmic plus cytosolic localisation (UniProt P22314 subcellular; HPA subcellular). A mitochondrial location is also annotated, but a fine granular chromogenic pattern alone cannot establish mitochondrial identity (UniProt P22314 subcellular; standard IHC practice). Compare compartments within intact cells and against an appropriate positive tissue control before calling a pattern abnormal (standard IHC practice). Document counterstain quality and section morphology, since poor nuclear boundaries can make cytoplasmic deposition appear nuclear (standard IHC practice).
Could isoforms or epitope changes explain discordant UBA1 IHC patterns?
UBA1 has 2 annotated isoforms, but the supplied antibody evidence does not identify which epitopes this antibody recognizes (UniProt P22314 isoforms; caption M02810). The protein has annotated modifications, including phosphorylation near its amino terminus, without evidence here that they alter this antibody’s staining (UniProt P22314 modified residues; caption M02810). Do not assign a staining pattern to one isoform from chromogenic IHC alone (standard IHC practice). Compare the same antibody across matched sections and review any available epitope documentation before proposing an isoform-specific explanation (standard IHC practice). If results remain discordant, corroborate the cellular pattern with an independently characterized antibody or orthogonal assay (standard IHC practice).
How can IF help assess a questionable UBA1 IHC pattern?
Use IF as a separate corroborating experiment, since the selected antibody evidence describes paraffin-section chromogenic IHC rather than an IF protocol (caption M02810). Multiplex UBA1 with a marker identifying the expected cells, such as kidney tubular cells, to check cellular assignment (HPA tissue IHC; standard IF practice). Choose fluorophores with emission distinguishable from tissue autofluorescence and include single-channel controls when evaluating overlap (standard IF practice). UBA1 lacks a transmembrane segment and has nuclear and cytosolic locations; permeabilise sufficiently to reach intracellular epitopes, while checking that treatment preserves cell morphology (UniProt P22314 topology and subcellular; standard IF practice). Use a nuclear counterstain to assess nucleoplasmic enrichment (HPA subcellular; standard IF practice).
What should I check when UBA1 DAB staining looks diffuse?
First inspect a no-primary control for nonspecific secondary or detection signal and examine morphology for pigment or damaged tissue (standard IHC practice). The selected image used a peroxidase-linked secondary and DAB detection, so verify peroxidase blocking when diffuse brown signal persists (caption M02810; standard IHC practice). Its 10% goat-serum block and 1:50 primary incubation provide documented starting conditions for that image (caption M02810). Compare background in intact cells with tissue edges and necrotic regions before changing antibody concentration (standard IHC practice). Recheck wash consistency and reagent compatibility, then judge changes against a positive control processed in the same run (standard IHC practice).
How should I score UBA1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear versus cytoplasmic compartment before scoring, since UBA1 can occupy both locations (UniProt P22314 subcellular; HPA subcellular). An H-score can combine the percentage of positive cells with staining intensity, while percentage positive alone answers a simpler prevalence question (standard IHC practice). For spatial questions, report positive-cell density per mm² of evaluable tissue and specify excluded necrotic or damaged areas (standard IHC practice). Normalise each result to the relevant counted cells or evaluable tissue area, and apply one scoring threshold across matched runs (standard IHC practice). Include a consistent positive control because tissue expression and staining consistency vary across the supplied evidence (HPA tissue IHC).
How can I distinguish true UBA1 positivity from an artefact?
Give most weight to staining in intact nuclei, while assessing cytoplasmic signal separately because both compartments are supported for UBA1 (HPA tissue IHC; HPA subcellular). Check whether positive cells fit the sampled tissue: kidney tubular cells show high staining, whereas heart cardiomyocytes are listed as low (HPA tissue IHC). Strong signal confined to cut edges, necrosis, or tissue folds warrants caution and comparison with well-preserved regions (standard IHC practice). If brown deposits remain in a no-primary control, investigate endogenous peroxidase and detection background before attributing them to UBA1 (standard IHC practice). Interpret unusual compartment patterns only after reviewing morphology, controls, and the antibody’s staining conditions (caption M02810; standard IHC practice).
Boster reagents
Best UBA1 / Ubiquitin-like modifier-activating enzyme 1 IHC Antibodies
M02810 has human paraffin-section IHC data (M02810 IHC captions). Both antibodies list IHC and ICC/IF for human, mouse, and rat; neither has an IF image (catalog applications, reactivity and image alts).
Real IHC data
Anti-UBA1/Ube1 Rabbit Monoclonal Antibody
Cat # M02810
M02810 lists IHC and ICC/IF for human, mouse, and rat (catalog applications/reactivity); its IHC captions document paraffin sections of human parotid acinic cell carcinoma and clear cell renal cell carcinoma (M02810 IHC captions). M02810-2 lists IHC and ICC/IF for human, mouse, and rat (catalog applications/reactivity), but has no supplied IHC or IF image caption (M02810-2 image alts).
Which to pick: For tissue IHC, choose M02810: its own paraffin-section caption documents EDTA pH 8 retrieval and a 1:50 primary dilution; the fixative is unreported (M02810 IHC caption). For IF/ICC, both rabbit monoclonals list the application, with 1:50 for M02810 and 1:50–1:200 for M02810-2; neither has an IF figure (catalog host, applications, IF dilutions and image alts). For cross-species work, both list human, mouse, and rat reactivity, while only M02810 provides a documented human paraffin-section IHC example (catalog reactivity; M02810 IHC captions; M02810-2 image alts).
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); cytoplasm and mitochondrion annotated (UniProt). 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: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Hippocampal neurons and myocytes show low staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Low tissue specificity (HPA RNA). 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.