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NEURL1 / E3 ubiquitin-protein ligase NEURL1 · IHC design guide
Design Immunohistochemistry for NEURL1
Plan chromogenic NEURL1 IHC in paraffin sections using brain as a positive control and the catalog antibody at 1:100–1:300 (HPA tissue IHC; datasheet A30740). Assess cytoplasmic CNS staining, including high Bergmann glial signal, alongside a negative control (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NEURL1 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 | Predominantly CNS cytoplasm; Bergmann glia also show membrane staining (HPA tissue IHC) | |
| Staining pattern | CNS glial cytoplasm; Bergmann glia also at the membrane (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30740) | |
| Positive control | Cerebellum+3 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A30740) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Lower in medulloblastoma than cerebellum (UniProt) | |
| Isoform / epitope | 2 isoforms; map the epitope to both (UniProt) |
Section 1
Recommended NEURL1 IHC & IF Protocols
The catalog antibody has a datasheet IHC-P protocol (datasheet A30740); one published conjunctival melanoma protocol provides an additional starting point (PMC6849808).
| Sample | Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30740) |
| Fixation | Image fixative and duration unreported (datasheet A30740); 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 8.0 (datasheet A30740); 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-NEURL1, 1:100 - 1:300 (datasheet A30740) |
| 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 | NEURL1-positive staining in bergmann glia - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in CNS. No signal in the no-primary control. |
💡Decision noteTry heat-mediated Tris-EDTA retrieval at pH 8.0 first for the catalog antibody (datasheet A30740). The published excerpt does not specify its retrieval conditions (PMC6849808).
Section 2
What Is the Expected NEURL1 Staining Pattern?
NEURL1 staining is expected mainly in the cytoplasm of CNS cells, with high cytoplasmic or membrane staining in cerebellar Bergmann glia and medium staining in cortical and hippocampal glia (HPA tissue IHC). Perinuclear, dendritic and postsynaptic localization is also described, chiefly from neuronal evidence by similarity (UniProt O76050). NEURL1 has no transmembrane segment (UniProt O76050 topology). HPA reports medium consistency between tissue staining and RNA data (HPA tissue IHC).
| Strong cytoplasmic or membrane signal in cerebellar Bergmann glia. | This matches the clearest supplied tissue observation: Bergmann glia stain at a high level (HPA tissue IHC). Judge the pattern in the identified cells, alongside slide controls (general IHC practice). |
| Moderate signal in cortical or hippocampal glia, or pancreatic exocrine cells. | These are reported at medium levels (HPA tissue IHC). Signal outside the CNS can therefore be plausible; cell identity matters more than a whole-tissue positive call (HPA tissue IHC). |
| Predominantly nuclear staining with little cytoplasmic signal. | This conflicts with the reported cytoplasmic CNS profile and UniProt localization (HPA tissue IHC; UniProt O76050). Treat it as a possible artefact and review controls before scoring it as NEURL1. |
| Strong staining in a cell type reported as undetected, such as adipocytes. | Adipocytes are reported as undetected in adipose tissue (HPA tissue IHC). Check cell identification and controls; cross-reactivity or endogenous detection activity are possible explanations (general IHC practice), not established causes. |
| Widespread weak colour across cells and tissue spaces. | This is difficult to assign to the cell-specific HPA pattern (HPA tissue IHC). Compare the negative detection control and inspect tissue edges and deposits before interpreting it as expression (general IHC practice). |
💡Expected NEURL1 appearanceCall a section positive when Bergmann glia show high cytoplasmic or membrane staining, or reported glial populations show medium staining (HPA tissue IHC); dominant nuclear colour or uniform haze is suspect against that pattern (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Cerebellar Bergmann glia provide the strongest reported tissue pattern; cortical and hippocampal glia and pancreatic exocrine cells are medium, while caudate glia are low (HPA tissue IHC). |
| Reported subcellular distribution | HPA describes mainly cytoplasmic CNS expression, including cytoplasmic or membrane staining in Bergmann glia (HPA tissue IHC). UniProt also lists perinuclear, dendritic and postsynaptic sites (UniProt O76050). |
| Antibody evidence and limits | The listed antibody, HPA044204, has IHC Enhanced validation; the tissue profile has medium staining-to-RNA consistency (HPA antibodies; HPA tissue IHC). These ratings support interpretation but do not establish every stained cell's identity. |
| Protein forms | UniProt lists 2 isoforms, no signal peptide or propeptide, and a chain spanning residues 2–574 (UniProt O76050). Without an antibody epitope, the supplied record cannot predict isoform-specific staining. |
| RNA and protein discordance | UniProt reports expression in bone marrow, while HPA reports hematopoietic cells there as undetected by tissue IHC (UniProt O76050; HPA tissue IHC). Do not treat the expression report as proof of visible IHC signal. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in cerebellar Bergmann glia. | The result misses HPA's high-staining reference population (HPA tissue IHC); the supplied sources do not identify a target-specific fixation cause. | Confirm the cell population, then check positive-control performance, antibody dilution, retrieval and detection using the assay instructions (general IHC practice). |
| Only faint staining in caudate glia. | Low staining is reported for this population (HPA tissue IHC). | Use the reported high-staining Bergmann glia to judge whether the run worked; avoid calling a low-staining caudate section an assay failure on its own (HPA tissue IHC). |
| Strong, predominantly nuclear colour. | Nuclear dominance does not match the supplied NEURL1 localization (HPA tissue IHC; UniProt O76050). | Review morphology and the negative detection control; inspect counterstain and chromogen deposits before assigning target signal (general IHC practice). |
| Adipocytes or other reported negative cells stain strongly. | Adipocytes are undetected in the HPA tissue profile (HPA tissue IHC); cross-reactivity or endogenous activity remains possible (general IHC practice). | Verify cell identity and compare appropriate negative controls. Reassess blocking or detection conditions if controls show the same colour (general IHC practice). |
| Diffuse colour obscures cell boundaries. | Background can prevent assessment of the cell-specific HPA pattern (HPA tissue IHC); its source cannot be determined from appearance alone. | Compare the negative detection control and review blocking, washes and chromogen development under the assay instructions (general IHC practice). |
| Can IF/ICC confirm the same compartment? | HPA provides no main ICC-IF location or cell-line images for NEURL1 (HPA subcellular record). | Treat IF/ICC localization as unconfirmed by this HPA record; use the separate IF/ICC guide for its assay design, and do not transfer an IHC staining call directly to IF. |
Experiment evidence
IHC & IF images showing NEURL1 staining
A30740 · IHCImmunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100(4° overnight). High-pressure and temperature Tr
A11177 · IHCImmunohistochemistry (IHC) analyzes of Neuralized-1 (V250) pAb in paraffin-embedded human breast carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107954-NEURL1/tissue/cerebral+cortex#imid_11842239
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000107954-NEURL1/tissue/cerebellum#imid_11842250
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107954-NEURL1/tissue/colon#imid_11842165
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107954-NEURL1/tissue/liver#imid_11842174
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NEURL1 IHC & IF
🧪Run cerebellum first and assess staining in Bergmann glia, which show High cytoplasmic/membrane signal (HPA: Cerebellum, Bergmann glia). Use adipose tissue as a negative, with adipocytes reported as Not detected (HPA: Adipose tissue, adipocytes); on the cerebellar slide, cells outside the stained Bergmann glia should retain counterstain without specific chromogen, but treat them as internal negatives only if they lack staining relative to the no-primary control.
Positive control tissue: Cerebellum (Bergmann glia - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for NEURL1; derive a cell-line control from the positive tissue's cell type (Bergmann glia - cytoplasm/membrane) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control and an isotype control matched to the primary antibody’s host species and immunoglobulin class; confirm specificity with NEURL1 knockout tissue or peptide preabsorption, as used for the catalog antibody’s brain IHC image (caption: immunogen-peptide preabsorption). For chromogenic cerebellar IHC, quench endogenous peroxidase and inspect the control slide for residual background before scoring.
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A30740 paraffin-section caption does not state a fixative (caption: fixative unreported). The caption reports high-pressure, high-temperature Tris-EDTA retrieval at pH 8.0 and antibody dilution 1:100 overnight at 4 °C; it supports that retrieval protocol, but does not establish whether retrieval is required (caption: human brain IHC). The evidence does not establish that frozen sections or IF are easier; for brain IF, check autofluorescence with a no-primary control, while paraffin IHC requires attention to retrieval-related background.
HPA tissue IHC evidence for NEURL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Cerebellum | Bergmann glia - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NEURL1 IHC Tips
Troubleshoot NEURL1 staining in paraffin section chromogenic IHC; the IF entry addresses planning a separate fluorescence assay.
Which retrieval conditions should I start with for weak NEURL1 staining?
Start with heat-mediated Tris-EDTA retrieval at pH 8.0 for paraffin sections (datasheet A30740). The selected human brain image used high-pressure, high-temperature retrieval in that buffer, followed by antibody at 1:100 overnight at 4 °C (caption A30740). If signal is weak, vary heating duration in a small pilot while holding the buffer, antibody dilution, and detection conditions constant (standard IHC practice). Compare signal in Bergmann glia with background in adjacent structures, because cytoplasmic or membrane staining there is a documented positive pattern (HPA: cerebellum). Record tissue damage alongside signal; stronger staining alone does not establish specific detection (standard IHC practice).
Could fixation explain weak or uneven NEURL1 staining?
NEURL1-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A30740). Record the actual fixative and fixation interval for each specimen, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Excessive or uneven fixation can change epitope accessibility in IHC generally, so investigate that possibility with matched processing rather than assigning it a NEURL1-specific effect (standard IHC practice). The published image conditions support Tris-EDTA retrieval at pH 8.0 and antibody incubation at 1:100 overnight at 4 °C; they do not establish an optimal fixation duration (caption A30740).
Where should convincing NEURL1 chromogen appear in brain sections?
Assess cytoplasmic and perinuclear signal first, while allowing membrane and dendritic staining where cell morphology supports it (UniProt O76050 subcellular location). NEURL1 has no transmembrane segment, so membrane-associated staining should not be interpreted as proof of a membrane-spanning protein (UniProt O76050 topology). In cerebellum, Bergmann glia show high cytoplasmic or membrane staining; glial cells show medium staining in cerebral cortex and hippocampus (HPA: tissue IHC). Neuronal cell bodies, apical dendrites, and postsynaptic sites are also reported locations, partly by similarity (UniProt O76050 subcellular location). Score cell identity and compartment separately, especially when a dense neuropil obscures cell boundaries (standard IHC practice).
Can I interpret a negative section as loss of every NEURL1 isoform?
No: NEURL1 has 2 listed isoforms, and the supplied evidence does not map the catalog antibody’s epitope to either one (UniProt O76050 isoforms; caption A30740). Its NHR domains span residues 61–217 and 292–447, but domain location alone does not establish which isoform the antibody detects (UniProt O76050 domains; standard antibody interpretation). Treat a negative section as absence of detectable staining under the tested conditions, then check tissue preservation, retrieval, and a positive tissue control (standard IHC practice). Peptide pre-absorption reduced staining in the selected image, supporting peptide-dependent signal without resolving isoform coverage or excluding every form of cross-reactivity (caption A30740; standard antibody interpretation).
How should I plan a separate NEURL1 immunofluorescence experiment?
Use the paraffin-section IHC result as a starting reference, not as IF validation: the selected image documents chromogenic tissue staining, while no ICC/IF images are listed in the supplied HPA record (caption A30740; HPA: subcellular). For a 2-color pilot, pair NEURL1 with a validated marker of the expected cell type, such as a glial marker when examining Bergmann glia (HPA: cerebellum; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence and include single-color controls for bleed-through assessment (standard IF practice). Because NEURL1 is reported in cytoplasmic, perinuclear, membrane, and dendritic locations without a transmembrane segment, optimize permeabilisation for the epitope’s accessible side once its location is established (UniProt O76050 subcellular location and topology; standard IF practice).
How can I reduce diffuse or misleading NEURL1 background?
First compare no-primary and full-stain sections, and inspect pigment, tissue folds, edges, and necrotic regions separately from intact cells (standard IHC practice). Block endogenous peroxidase before chromogenic detection and control nonspecific binding with an appropriate protein block and washes (standard IHC practice). If background remains high, titrate around the documented 1:100 overnight, 4 °C incubation while keeping Tris-EDTA retrieval at pH 8.0 fixed (caption A30740; standard IHC practice). The selected image includes a peptide-preabsorbed negative control; use that result as evidence for peptide-dependent staining, while checking whether residual signal follows tissue edges or endogenous enzyme activity (caption A30740; standard IHC interpretation).
What is a defensible way to quantify NEURL1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Choose one prespecified readout: percentage of positive cells, staining density per mm², or an H-score combining intensity and positive-cell fraction (standard IHC scoring practice). Segment or annotate the same cell population and anatomical region across sections, then normalise counts to eligible cells or measured tissue area rather than total image area (standard IHC scoring practice). Report cytoplasmic and membrane-associated signal separately when boundaries permit, because both occur in Bergmann glia (HPA: cerebellum). Keep retrieval, antibody incubation, chromogen development, imaging, and threshold settings consistent; the selected example used 1:100 overnight at 4 °C after pH 8.0 retrieval (caption A30740; standard IHC scoring practice).
How do I distinguish true NEURL1 staining from artefact?
A plausible positive follows an expected cell and compartment: Bergmann glial cytoplasm or membrane in cerebellum, or glial cells in cortex and hippocampus (HPA: tissue IHC). Perinuclear or dendritic signal can also fit the reported localisation, but morphology alone cannot prove target identity (UniProt O76050 subcellular location; standard IHC interpretation). Question staining confined to section edges, folds, necrosis, or sites of endogenous peroxidase activity, and compare no-primary and peptide-competition controls where available (standard IHC practice; caption A30740). Be cautious with adipocytes, which were not detected in the supplied adipose-tissue profile, and with broad claims of validation: HPA rates tissue staining as Enhanced while describing only medium agreement with RNA expression (HPA: tissue IHC).
Boster reagents
Best NEURL1 / E3 ubiquitin-protein ligase NEURL1 IHC Antibodies
Two anti-NEURL1 antibodies have paraffin-section IHC images from human tissue (catalog image captions). Both list human, mouse, and rat reactivity; A30740 also lists IF/ICC applications (catalog applications/reactivity).
Real IHC data
Anti-NEURL1 Antibody
Cat # A30740
Real IHC data
Anti-Neuralized-1 (V250) NEURL1 Antibody
Cat # A11177
A30740 shows IHC in paraffin-embedded human brain at 1:100, with Tris-EDTA pH 8.0 retrieval and a peptide-preabsorption control (A30740 image caption). A11177 shows IHC in paraffin-embedded human breast carcinoma at 1:50 (A11177 image caption).
Which to pick: Choose A30740 for tissue IHC when its documented retrieval and peptide-preabsorption control are useful; its image shows paraffin-embedded human brain, but the fixative is unreported (A30740 image caption). Choose A30740 for IF/ICC because those applications are listed, although the payload provides no IF image (A30740 catalog applications/images). Both antibodies are rabbit polyclonals listing human, mouse, and rat reactivity; their IHC images document human tissue only, and A11177 offers a paraffin-embedded breast carcinoma example with fixative unreported (catalog host/type/reactivity; A11177 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O76050 (NEUL1_HUMAN, E3 ubiquitin-protein ligase NEURL1).
- Human Protein Atlas. NEURL1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NEURL1 subcellular location (ICC-IF): Highest expression in SiHa: 10.7 nTPM.
- Human Protein Atlas. NEURL1 antibody validation summary (1 antibodies).
- Conjunctival melanoma copy number alterations and correlation with mutation status, tumor features, and clinical outcome. Pigment cell & melanoma research 2019 — PMC6849808.
- PubMed PMID:9519875 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NEURL1 (E3 ubiquitin-protein ligase NEURL1). Expected localisation Predominantly CNS cytoplasm; Bergmann glia also show membrane staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30740). Validated IHC antibody: A30740 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Bergmann glia - cytoplasm/membrane); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human brain tissue; fixative not specified (datasheet A30740); 2. Fixation: Image fixative and duration unreported (datasheet A30740); 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 8.0 (datasheet A30740); 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-NEURL1, 1:100 - 1:300 (datasheet A30740); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A30740); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; map the epitope to both (UniProt); Regulation: Lower in medulloblastoma than cerebellum (UniProt). Evidence: UniProt O76050, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in SiHa: 10.7 nTPM, Datasheet A30740, PMC6849808. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NEURL1/immunohistochemistry-ihc-guide.