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MAEA / E3 ubiquitin-protein transferase MAEA · IHC design guide
Design Immunohistochemistry for MAEA
MAEA IHC shows general nuclear and cytoplasmic tissue staining, with high signal in colon glandular cells and bronchial respiratory epithelial cells (HPA tissue IHC). This guide pairs those expectations with the catalog antibody’s 1:100–1:300 IHC dilution range (datasheet A08766-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAEA 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | High glandular and epithelial staining; nuclear and cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A08766-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 5 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended MAEA IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A08766-1) with two published MAEA IHC workflows (PMC12450695; PMC10506284).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A08766-1) |
| Fixation | Image fixative and duration unreported (datasheet A08766-1); 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 A08766-1); 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-MAEA, 1:100 - 1:300 (datasheet A08766-1) |
| 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 | MAEA-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 9.0 for the catalog antibody (datasheet: A08766-1); the osteosarcoma paper does not specify its retrieval conditions (PMC12450695).
Section 2
What Is the Expected MAEA Staining Pattern?
MAEA staining should be assessed in nuclei and cytoplasm across many cell types; high staining is reported in selected epithelial and neuronal cells (HPA: general nuclear and cytoplasmic expression; Low tissue specificity). Macrophages may also show cell-surface staining that varies with maturation (UniProt Q7L5Y9: macrophage localisation). The tissue IHC profile is Approved, with medium consistency between staining and RNA data (HPA: reliability). MAEA has no transmembrane segment (UniProt Q7L5Y9: topology).
| Distinct nuclear and cytoplasmic staining in colon glandular cells or bronchial respiratory epithelial cells. | This fits the reported compartment pattern and high staining in those cells (HPA: general nuclear and cytoplasmic expression; High in colon and bronchus). Judge intensity against cells in the same section; HPA's Approved rating has medium consistency with RNA data (HPA: reliability). |
| Signal confined to an unexpected compartment, with no convincing nuclear or cytoplasmic staining. | Check for an artefact before treating that distribution as MAEA: the tissue profile is nuclear and cytoplasmic (HPA: tissue IHC). Cell-surface staining can be plausible in macrophages, particularly mature ones (UniProt Q7L5Y9: macrophage localisation). |
| Strong staining in a cell population outside the expected high-staining populations. | Compare its identity with the tissue profile and controls; cross-reactivity or endogenous detection activity is possible (general IHC practice). An unlisted cell type is not automatically negative because MAEA is ubiquitous (UniProt Q7L5Y9: tissue specificity). |
| Diffuse colour over tissue and empty areas, obscuring cell boundaries. | Treat this as background until controls and a cleaner run establish a cellular pattern (general IHC practice). It cannot be scored reliably against HPA's nuclear and cytoplasmic profile (HPA: tissue IHC). |
| No signal in colon glandular cells or cerebellar Purkinje cells. | Investigate the run: both are reported as High, though the HPA profile has medium consistency with RNA data (HPA: tissue IHC; reliability). A single negative section does not establish MAEA absence (general IHC practice). |
💡Expected MAEA appearanceA convincing positive shows discernible nuclear and cytoplasmic staining in reported High cells, such as colon glandular cells (HPA: tissue IHC); diffuse colour without cell-specific localisation suggests background (general IHC practice).
| Tissue and cell selection | Colon glandular cells and cerebellar Purkinje cells are reported High; liver hepatocytes and soft-tissue fibroblasts are Low (HPA: tissue IHC). Use the latter as lower-signal comparators, not proven negatives; HPA lists no negative population (HPA: tissue IHC). |
| Macrophage maturation and compartment | Immature macrophages are predominantly nuclear, whereas mature macrophages are predominantly cell-surface positive (UniProt Q7L5Y9: subcellular location). Interpret a macrophage membrane-like outline in that context; MAEA has no transmembrane segment (UniProt Q7L5Y9: topology). |
| Antibody evidence | HPA058185 has Approved IHC status; HPA036886 has no listed IHC status (HPA: antibodies). Those ratings are antibody-specific. The overall tissue profile is Approved with medium RNA–staining consistency (HPA: reliability). |
| Isoforms and processing | Five isoforms are listed, with no signal peptide, propeptide or processed chain reported (UniProt Q7L5Y9: isoforms; processing). These facts do not identify which isoforms an antibody detects or predict loss of staining after tissue preparation. |
| Target-specific fixation sensitivity | The supplied HPA staining profile and UniProt record provide no MAEA-specific fixation effect (HPA: tissue IHC; UniProt Q7L5Y9). Assess retrieval empirically as a general IHC workflow variable; do not infer its effect from tissue staining levels (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High tissue has no discernible nuclear or cytoplasmic signal. | A failed staining or detection step is possible (general IHC practice); colon glandular cells are reported High (HPA: tissue IHC). | Check the run's positive control and detection reagents; review retrieval and primary-antibody conditions as general IHC variables (general IHC practice). Do not assign an MAEA-specific fixation cause from these sources. |
| Colour spreads across the section or appears where no cells are present. | Nonspecific binding, incomplete washing or detection-system background can obscure localisation (general IHC practice). | Compare with a primary-omission control, then review blocking, washing and detection conditions (general IHC practice). Score MAEA only when cellular staining is resolved. |
| Signal appears mainly at macrophage surfaces. | A surface-associated pattern is reported for mature macrophages (UniProt Q7L5Y9: macrophage localisation). | Confirm macrophage identity and compare nuclear and surface distributions before calling the outline artefactual (general IHC practice; UniProt Q7L5Y9: macrophage localisation). |
| Unexpected cells stain strongly while reported High cells do not. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); MAEA is also described as ubiquitous (UniProt Q7L5Y9: tissue specificity). | Check cell identity and primary-omission controls; compare with reported High populations on a separate section (general IHC practice; HPA: tissue IHC). Avoid declaring every unlisted cell negative. |
| Liver hepatocytes show weak staining. | Low staining is reported for hepatocytes (HPA: tissue IHC). Weak signal alone does not demonstrate assay failure. | Interpret alongside a reported High population and the run's controls before changing staining conditions (HPA: tissue IHC; general IHC practice). |
| IF/ICC shows mostly nucleoplasmic signal; is that consistent with MAEA? | Nucleoplasm is the enhanced main location in HPA ICC-IF images (HPA: subcellular); tissue IHC has a broader nuclear and cytoplasmic profile (HPA: tissue IHC). | Interpret IF/ICC on its own guide page; HPA lists ICC as Supported for HPA036886 and HPA058185 (HPA: antibodies). Do not use the IF pattern alone to reject a tissue IHC result. |
Experiment evidence
IHC & IF images showing MAEA staining
A08766-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was used
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000090316-MAEA/tissue/cerebral+cortex#imid_20097189
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000090316-MAEA/tissue/colon#imid_20097115
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000090316-MAEA/tissue/liver#imid_20097124
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000090316-MAEA/tissue/kidney#imid_20097109
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036886; green = MAEA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036886; green = MAEA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAEA IHC & IF
🧪Run breast first and score glandular cells, which show high MAEA staining (HPA: High in breast glandular cells). HPA detects MAEA in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells without visible staining on the breast slide as internal background references only, not confirmed MAEA-negative cells (HPA: no negative rows; MAEA detected in all 45 scored tissues).
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: None in HPA: MAEA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAEA in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Run secondary-only and host-species- and clonality-matched isotype controls, with MAEA knockout material or a validated peptide-blocking control as a biological specificity check (standard IHC practice). For chromogenic detection on breast sections, quench endogenous peroxidase and assess endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (A08766-1 caption: fixative unreported). The caption uses Tris-EDTA at pH 9.0 for retrieval and primary antibody at 1:200, but does not establish that retrieval is required (A08766-1 caption). Whether frozen sections or IF/ICC would be easier is unreported; breast stroma may add autofluorescent background in IF (standard IF practice).
HPA tissue IHC evidence for MAEA
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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MAEA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MAEA IHC Tips
These questions focus on interpreting MAEA staining in paraffin sections using chromogenic IHC; the immunofluorescence entry addresses the separate IF workflow.
Which retrieval conditions should I try first for weak MAEA staining?
Start with heat-mediated Tris-EDTA, pH 9.0, antigen retrieval for paraffin sections (datasheet A08766-1). The selected tonsil image used this buffer and the catalog antibody at 1:200 after an overnight incubation at 4°C, but its caption does not report fixation or heating duration (A08766-1 caption). If staining remains weak, vary heating time in a controlled series while keeping antibody incubation and detection constant; include a positive section and a no-primary control (standard IHC practice). Compare tissue morphology and nuclear and cytoplasmic signal across conditions, since excessive heating can damage sections and increase diffuse background (standard IHC practice; HPA: general nuclear and cytoplasmic expression).
How should I assess whether fixation is affecting MAEA IHC?
The tonsil caption identifies paraffin sections but does not state a fixative, so MAEA-specific fixation sensitivity is unknown (A08766-1 caption). Document the fixative and fixation interval for each specimen, then compare matched sections processed under the same retrieval and detection conditions (standard IHC practice). If staining differs between batches, test a controlled fixation series with the same positive reference and assess nuclear, cytoplasmic, and background signal separately, without assigning the change to a MAEA-specific mechanism (standard IHC practice; HPA: general nuclear and cytoplasmic expression). Record morphology and processing history before changing antibody concentration, because damaged architecture can confound compartment scoring (standard IHC practice).
Should MAEA staining be nuclear, cytoplasmic, or membranous?
Expect nuclear and cytoplasmic staining across tissues, and evaluate each compartment separately (HPA: general nuclear and cytoplasmic expression). MAEA is also annotated in the nucleoplasm, nuclear matrix, cytoskeleton, and cell membrane, with macrophage membrane detection reported at protein level (UniProt Q7L5Y9: subcellular location and tissue specificity). A nucleoplasmic pattern has independent IF support, while the described macrophage shift from predominantly nuclear in immature cells to surface enriched in mature cells is annotated by similarity (HPA: nucleoplasm enhanced; UniProt Q7L5Y9: macrophage localisation). Interpret apparent membrane staining against cell boundaries and matched controls, because MAEA has no transmembrane segment and a rim alone does not establish surface localisation (UniProt Q7L5Y9: topology; standard IHC practice).
Could MAEA isoforms or epitope masking explain inconsistent staining?
MAEA has 5 annotated isoforms, but the supplied caption does not identify this antibody’s epitope or establish which isoforms it recognizes (UniProt Q7L5Y9: isoforms; A08766-1 caption). Its LisH and CTLH regions span residues 121–153 and 159–216, respectively, so map any subsequently documented immunogen against the relevant isoform sequences before assigning staining differences to splice variation (UniProt Q7L5Y9: domains and isoforms). Phosphothreonine at residue 28 is annotated, but no supplied evidence links that modification to this antibody’s IHC performance (UniProt Q7L5Y9: modified residues; A08766-1 caption). Compare retrieval conditions and controls first, then seek epitope and isoform validation before making an isoform-specific claim (standard IHC practice).
How can IF help resolve ambiguous MAEA localisation in IHC?
For a separate IF experiment, multiplex MAEA with a macrophage marker such as CD68 when assessing macrophages, and inspect cell identity alongside nuclear and surface signal (UniProt Q7L5Y9: macrophage localisation; standard IF practice). Choose spectrally separated fluorophores, favoring a far-red channel when tissue autofluorescence obscures shorter wavelengths, and include single-stain controls (standard IF practice). Use permeabilisation for an intracellular epitope, but assess a proposed surface epitope without permeabilisation first; the antibody epitope is unspecified and MAEA has no transmembrane segment (A08766-1 caption; UniProt Q7L5Y9: topology; standard IF practice). HPA reports enhanced nucleoplasmic localisation from ICC/IF images, which can guide comparison with IHC but does not establish this catalog antibody’s IF fixation or permeabilisation conditions (HPA: subcellular summary; A08766-1 caption).
How do I reduce diffuse or misleading MAEA chromogen signal?
First compare a no-primary control, a tissue reference, and the stained section to distinguish detection background from plausible nuclear and cytoplasmic MAEA signal (standard IHC practice; HPA: general nuclear and cytoplasmic expression). Check endogenous peroxidase blocking before DAB development and assess nonspecific secondary binding with appropriate controls; these are general chromogenic workflow steps, not MAEA-specific findings (standard IHC practice). The selected tonsil caption reports primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature (A08766-1 caption). If background persists, titrate antibody and detection time while holding retrieval and control sections constant, and exclude tissue folds or damaged areas from assessment (standard IHC practice).
How should I score MAEA staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA describes general nuclear and cytoplasmic expression and high staining in several epithelial and neuronal populations (HPA: tissue IHC profile). For a cell-based readout, record percent positive cells and intensity, then calculate a compartment-specific H-score from intensity-weighted percentages; keep thresholds and exposure conditions fixed across samples (standard IHC practice). Where cell numbers vary, report positive-cell density per mm² of viable tissue or normalize positive cells to all eligible cells of the same type (standard IHC practice). Exclude folds, edges, and necrotic regions, and report nuclear and cytoplasmic scores separately rather than combining biologically different patterns (standard IHC practice; HPA: tissue IHC profile).
What distinguishes credible MAEA staining from an IHC artefact?
Credible staining should occur in intact cells, track recognizable nuclei or cytoplasm, and exceed the matched no-primary control (HPA: general nuclear and cytoplasmic expression; standard IHC practice). High signal in breast glandular cells or bronchial respiratory epithelial cells and lower signal in hepatocytes can provide contextual checks, though HPA labels its tissue evidence Approved with medium RNA consistency (HPA: tissue IHC). Examine a proposed macrophage surface rim against cell boundaries, since membrane detection is reported but MAEA lacks a transmembrane segment (UniProt Q7L5Y9: tissue specificity and topology). Discount isolated edge staining, necrotic deposits, and signal reproduced by the no-primary or peroxidase control before treating a compartment or cell type as positive (standard IHC practice).
Boster reagents
Best MAEA / E3 ubiquitin-protein transferase MAEA IHC Antibodies
A08766-1 has an IHC figure from paraffin-embedded human tonsil (catalog image caption). IF/ICC are listed applications without an IF figure (catalog applications and image inventory); reactivity lists human, mouse and rat (catalog reactivity).
Real IHC data
Anti-MAEA/Emp Antibody
Cat # A08766-1
A08766-1 is shown by IHC on paraffin-embedded human tonsil (catalog image caption). A08766-1 also lists IF/ICC applications and human, mouse and rat reactivity, but has no IF image (catalog applications, reactivity and image inventory).
Which to pick: For tissue IHC, choose A08766-1: its figure documents paraffin-embedded human tonsil, Tris-EDTA retrieval at pH 9.0, and primary antibody at 1:200 overnight at 4 °C (catalog IHC image caption). For IF/ICC, A08766-1 lists those applications and an IF dilution of 1:50, but has no IF figure (catalog applications, IF dilution and image inventory). For cross-species work, A08766-1 is polyclonal and lists human, mouse and rat reactivity (catalog dilution data and reactivity); its IHC figure shows only human tonsil, and the fixative is unreported (catalog IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q7L5Y9 (MAEA_HUMAN, E3 ubiquitin-protein transferase MAEA).
- Human Protein Atlas. MAEA tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MAEA subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. MAEA antibody validation summary (2 antibodies).
- Identification of MAEA protein as a potential target for chemoresistance in osteosarcoma using bioinformatics and proteomic analysis. Frontiers in oncology 2025 — PMC12450695.
- Plasma biomarkers for prediction of early tumor recurrence after resection of pancreatic ductal adenocarcinoma. Scientific reports 2021 — PMC8021576.
- The E3 ubiquitin ligase MAEA promotes macrophage phagocytosis and inhibits gastrointestinal cancer progression by mediating PARP1 ubiquitination and degradation. International journal of biological sciences 2025 — PMC11844278.
- LAD1 promotes malignant progression by diminishing ubiquitin-dependent degradation of vimentin in gastric cancer. Journal of translational medicine 2023 — PMC10506284.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:9763581 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAEA (E3 ubiquitin-protein transferase MAEA). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A08766-1). Validated IHC antibody: A08766-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A08766-1); 2. Fixation: Image fixative and duration unreported (datasheet A08766-1); 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 A08766-1); 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-MAEA, 1:100 - 1:300 (datasheet A08766-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; epitope coverage is unspecified (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q7L5Y9, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A08766-1, PMC12450695, PMC10506284. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAEA/immunohistochemistry-ihc-guide.