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ELAVL4 / ELAV-like protein 4 · IHC design guide
Design Immunohistochemistry for ELAVL4
Plan ELAVL4 chromogenic IHC in paraffin sections using cytoplasmic staining in brain, adrenal medulla and spermatogonia as a reference (HPA tissue IHC). This guide covers fixation consistency, controls and interpretation of cell-specific staining.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ELAVL4 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 | Cytoplasmic in brain, adrenal medulla and spermatogonia (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in neurons, medullary cells and spermatogonia (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9698) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 PB9698) | |
| Caveat | Staining varies by cell type within a section (HPA tissue IHC) | |
| Regulation | Brain and adrenal gland enriched RNA (HPA RNA) | |
| Isoform / epitope | Six isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended ELAVL4 IHC & IF Protocols
The catalog antibody has an IHC-P protocol; the published options below provide chromogenic ELAVL4 staining methods for human spinal cord and small cell lung cancer sections (PMC6613039; PMC5152694).
| Sample | Paraffin-embedded mouse brain tissues; fixative not specified (datasheet PB9698) |
| Fixation | Image fixative and duration unreported (datasheet PB9698); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet PB9698) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9698) |
| Primary antibody | Rabbit anti-ELAVL4, 0.5-1μg/ml (datasheet PB9698) |
| Primary incubation | Overnight at 4 °C (datasheet PB9698) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9698) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ELAVL4-positive staining in medullary cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in brain, adrenal medulla, and spermatogonia in seminiferous ducts. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet: PB9698); the spinal cord protocol also reports citrate pH 6.0 retrieval (PMC6613039).
Section 2
What Is the Expected ELAVL4 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic ELAVL4 staining in brain neurons, adrenal medullary cells and testicular spermatogonia (HPA tissue IHC: Enhanced reliability; High in these cells). Neuronal cell bodies and projections are plausible sites because ELAVL4 localizes to the perikaryon, dendrites and axons (UniProt P26378: subcellular location). ELAVL4 has no transmembrane segment, so a membrane-restricted pattern would be unexpected (UniProt P26378: topology).
| Strong cytoplasmic staining in neurons of cerebral cortex or hippocampus. | This matches the reported high neuronal staining in both regions (HPA tissue IHC: High in neuronal cells). Cytoplasm is the expected tissue-IHC compartment (HPA tissue IHC: cytoplasmic profile). Assess cells individually: a positive neuron need not make every surrounding cell positive. |
| Staining is mainly confined to nuclei or cell borders in a brain section. | Treat this as discordant with the cytoplasmic tissue-IHC pattern, and review controls before calling it ELAVL4 (HPA tissue IHC: cytoplasmic profile). Nuclear ICC-IF staining is separately reported and does not establish a nuclear paraffin-IHC pattern (HPA subcellular: approved nucleoplasm and vesicles). |
| Strong staining appears in adipocytes or bronchial respiratory epithelium. | These cell types are reported as not detected (HPA tissue IHC: adipocytes; bronchial respiratory epithelial cells). Consider antibody cross-reactivity or endogenous detection activity; compare the signal with the same run's controls (standard IHC practice). |
| Color spreads across many cell types or covers the section without clear cell boundaries. | Diffuse background is insufficient to identify ELAVL4-positive cells. Compare the distribution with high-staining neuronal or medullary cells (HPA tissue IHC: High). Check the no-primary control and detection reagents for nonspecific signal (standard IHC practice). |
| A cerebral cortex control shows no convincing neuronal stain. | Cerebral cortical neurons are reported as High, so a blank control makes the run difficult to interpret (HPA tissue IHC: cerebral cortex). Review section integrity, antigen retrieval, antibody dilution and detection performance before interpreting negative test tissue (standard IHC practice). |
💡Expected ELAVL4 appearanceCall a paraffin-IHC result positive when strong cytoplasmic staining identifies the expected neurons, adrenal medullary cells or spermatogonia; isolated strong signal in a reported negative cell type needs investigation (HPA tissue IHC: cytoplasmic profile; High positives; Not detected negatives).
| Tissue and cell selection | Cerebral cortex, hippocampus, caudate, adrenal medulla and testicular spermatogonia offer reported High staining; adipocytes offer a reported Not detected comparison (HPA tissue IHC). Score the named cell population, not the whole tissue. |
| Compartment and topology | Cytoplasmic tissue staining agrees with ELAVL4 in perikarya and neuronal projections (HPA tissue IHC: cytoplasmic profile; UniProt P26378: subcellular location). Its lack of a transmembrane segment gives no basis to expect an exclusive membrane outline (UniProt P26378: topology). |
| IHC antibody validation | The listed mouse monoclonal CAB004442 has Enhanced IHC status; the listed rabbit polyclonal HPA043047 has Approved ICC status, with no IHC status supplied (HPA antibodies). Keep validation claims tied to the application and antibody used. |
| What should IF/ICC show? | HPA reports approved nucleoplasm and vesicle localization in ICC-IF, whereas its tissue-IHC summary is cytoplasmic (HPA subcellular; HPA tissue IHC). Interpret the two applications on their own evidence; this section supplies no IF/ICC protocol. |
| Isoforms, epitopes and retrieval | UniProt lists 6 ELAVL4 isoforms and 3 RNA-recognition motifs (UniProt P26378: isoforms; domains). The supplied evidence does not map the IHC antibody epitope or establish an ELAVL4-specific retrieval condition; use documented antibody instructions and run controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive-control neurons are unstained. | A failed staining run is possible because cortical and hippocampal neurons are reported as High (HPA tissue IHC). | Verify tissue identity and section quality; then check retrieval, antibody preparation, dilution and detection steps against the assay instructions (standard IHC practice). |
| Most cells show pale, diffuse color. | Background or excessive detection signal may obscure the expected cell pattern (standard IHC practice). | Examine a no-primary control, review blocking and wash steps, and adjust detection or antibody concentration using the assay instructions (standard IHC practice). |
| Reported negative cells stain strongly. | Cross-reactivity or endogenous detection activity is possible; adipocytes and bronchial respiratory epithelium are Not detected in the HPA record (HPA tissue IHC). | Compare positive and negative tissue controls, include a no-primary control, and check the detection system's endogenous-activity blocking step (standard IHC practice). |
| Only nuclei stain in the paraffin section. | This conflicts with HPA's cytoplasmic tissue-IHC summary, although nucleoplasm is reported separately for ICC-IF (HPA tissue IHC; HPA subcellular). | Check counterstain and no-primary controls, then confirm that the antibody has IHC validation before assigning the nuclear signal to ELAVL4 (HPA antibodies; standard IHC practice). |
| Neuronal processes stain but cell bodies appear weak. | ELAVL4 is reported in both neuronal projections and the perikaryon; process-only staining warrants review rather than automatic rejection (UniProt P26378: subcellular location). | Inspect neuronal morphology and nearby positive cells at higher magnification; compare the pattern with the run's positive control (standard IHC practice). |
| Adrenal medulla stains but neighboring areas do not. | The reported High adrenal signal is assigned to medullary cells, so tissue-wide staining is not required (HPA tissue IHC: adrenal gland). | Identify medullary cells on the counterstained section and score their cytoplasmic signal separately from surrounding cells (HPA tissue IHC: cytoplasmic profile; standard IHC practice). |
Experiment evidence
IHC & IF images showing ELAVL4 staining
PB9698 · IHCIHC analysis of ELAVL4 using anti-ELAVL4 antibody (PB9698). ELAVL4 was detected in paraffin-embedded section of mouse brain tissues. Heat me
PB9698 · IHCIHC analysis of ELAVL4 using anti-ELAVL4 antibody (PB9698). ELAVL4 was detected in paraffin-embedded section of rat brain tissues. Heat medi
PB9698 · IFIF analysis of ELAVL4 using anti-ELAVL4 antibody (PB9698) ELAVL4 was detected in paraffin-embedded section of human glioma tissues. Heat med
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162374-ELAVL4/tissue/cerebral+cortex#imid_20813468
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000162374-ELAVL4/tissue/adrenal+gland#imid_1464
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162374-ELAVL4/tissue/colon#imid_20813386
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162374-ELAVL4/tissue/liver#imid_20813395
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA043047; green = ELAVL4, blue = nucleus, red = microtubules.
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA043047; green = ELAVL4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ELAVL4 IHC & IF
🧪Run cerebral cortex first and require staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes), and assess adjacent non-neuronal cells on the cortex slide as internal background without assuming they are ELAVL4-negative.
Positive control tissue: Adrenal gland (Medullary cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ELAVL4 in SH-SY5Y, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a rabbit IgG isotype control matched to the primary antibody’s clonality, and ELAVL4 knockout tissue if available to assess specificity (caption: rabbit anti-ELAVL4 PB9698; standard IHC practice). Block endogenous peroxidase and check endogenous biotin background in brain sections when using the caption’s biotin–streptavidin/DAB detection (caption: PB9698 detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected paraffin-section caption also leaves the fixative unreported (caption: PB9698). The reported mouse-brain IHC used citrate retrieval at pH 6 for 20 minutes, so verify retrieval with the catalog antibody in the intended specimens (caption: PB9698). The supplied evidence does not establish whether frozen sections or IF are easier; if adapting brain tissue to IF, assess lipofuscin autofluorescence (standard IF practice).
HPA tissue IHC evidence for ELAVL4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 | Medullary cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ELAVL4 IHC Tips
Troubleshoot ELAVL4 staining in paraffin sections by checking retrieval, cell identity, subcellular pattern, and matched controls before interpreting signal.
What retrieval should I try first when ELAVL4 staining is weak in paraffin sections?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet PB9698). The selected paraffin-section image used that retrieval before incubation with 1 μg/ml primary antibody overnight at 4°C (datasheet PB9698). Run a brain section alongside the test section so weak staining can be assessed against a tissue with reported neuronal expression (HPA: High in cerebral cortex neuronal cells). If signal remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Check whether stronger staining also raises background before adopting the change (standard IHC practice).
Could fixation explain absent or uneven ELAVL4 staining?
The selected paraffin-section caption does not state a fixative, so ELAVL4-specific fixation sensitivity is unknown for this antibody (datasheet PB9698). Record the fixative, fixation duration, section thickness, and processing history for both the test and positive-control sections (standard IHC practice). Process a brain control and test tissue together, then apply the same citrate retrieval at pH 6 for 20 minutes (datasheet PB9698; standard IHC practice). If staining differs, compare sections with documented processing histories before attributing the result to ELAVL4 abundance (standard IHC practice). Keep retrieval and detection settings matched during that comparison (standard IHC practice).
Where should convincing ELAVL4 signal appear in a chromogenic section?
Assess cellular context first: ELAVL4 is reported in neuronal cells of cerebral cortex and hippocampus and in adrenal medullary cells (HPA: High in those cells). Cytoplasmic staining is supported by tissue IHC, while the protein is also assigned to perikarya, dendrites, axons, and growth cones (HPA: cytoplasmic expression; UniProt P26378: subcellular location). Score cell bodies and identifiable processes separately when morphology permits, using a matched counterstain to locate nuclei (standard IHC practice). A nucleoplasmic signal is reported in cell imaging, so it merits review against the cytoplasmic tissue pattern rather than automatic rejection (HPA: approved nucleoplasm and vesicles; HPA: tissue IHC).
Could isoform choice or epitope accessibility change the IHC pattern?
ELAVL4 has 6 listed isoforms and 3 RNA recognition motifs, spanning residues 51–129, 137–217, and 302–380 (UniProt P26378: isoforms and domains). The supplied antibody caption does not identify its epitope, so the relative detection of those isoforms cannot be assigned from this IHC image (datasheet PB9698). Consult the antibody's epitope documentation before treating differences between specimens as isoform changes (standard IHC practice). If documentation remains unavailable, compare retrieval and staining on matched sections and report the result as total antibody-reactive ELAVL4 signal (standard IHC practice). Do not assign a phosphosite-specific pattern without a validated site-specific reagent (UniProt P26378: modified residues; standard IHC practice).
How should I plan a companion IF experiment to check the IHC pattern?
Pair ELAVL4 with a validated marker for the expected cell type, such as a neuronal marker when checking brain tissue, and inspect colocalisation within individual cells (HPA: High in brain neuronal cells; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before selecting the ELAVL4 channel (standard IF practice). ELAVL4 has no transmembrane segment and is assigned to intracellular compartments, so include a controlled permeabilisation step for access to its intracellular epitope (UniProt P26378: topology and subcellular location; standard IF practice). Include single-label and secondary-only controls to assess bleed-through and nonspecific fluorescence (standard IF practice). Interpret nuclear signal with care because cell imaging reports nucleoplasm and vesicles, whereas tissue IHC reports cytoplasmic expression (HPA: subcellular and tissue IHC).
How can I reduce diffuse or granular background without losing ELAVL4 signal?
Start by comparing the stained section with a no-primary control and an identically processed brain control (standard IHC practice; HPA: High in brain neuronal cells). The selected procedure used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, and biotin-based detection with DAB (datasheet PB9698). If background persists, check the peroxidase block and test a lower primary concentration or shorter chromogen development on adjacent sections (standard IHC practice). With biotin-based detection, assess endogenous biotin as a possible contributor to staining (datasheet PB9698: biotin-based detection; standard IHC practice). Preserve the same counterstain and imaging conditions when comparing adjustments (standard IHC practice).
What is a defensible way to quantify ELAVL4 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, then use the same segmentation and staining thresholds across sections (standard IHC practice). Report the percentage of positive target cells and staining intensity; an H-score can combine those measurements on a 0–300 scale (standard IHC practice). For cell-rich regions, positive-cell density per mm² is another option when the sampled area is recorded (standard IHC practice). Normalise counts to eligible cells or measured tissue area, and exclude folds, damaged edges, and necrotic regions consistently (standard IHC practice). Keep cytoplasmic and nuclear scores separate because reported tissue and cell-imaging patterns differ (HPA: tissue IHC and subcellular location).
How do I distinguish true ELAVL4 staining from artefact?
Look for reproducible staining in expected cells, including brain neuronal cells or adrenal medullary cells, with a plausible cytoplasmic component (HPA: High in those cells; HPA: cytoplasmic tissue expression). Compare those regions with adipocytes in adipose tissue, where ELAVL4 was not detected in the supplied tissue profile (HPA: Not detected in adipocytes). Review isolated nuclear-only staining cautiously: nucleoplasm is reported in cell imaging, while tissue IHC emphasises cytoplasm (HPA: approved nucleoplasm; HPA: tissue IHC). Discard edge staining and necrotic deposits from scoring, and use no-primary controls to reveal background (standard IHC practice). Check peroxidase blocking when DAB appears in unexpected cells (datasheet PB9698: DAB detection; standard IHC practice).
Boster reagents
Best ELAVL4 / ELAV-like protein 4 IHC Antibodies
PB9698 has real IHC images from paraffin sections of mouse and rat brain and human meningeoma, plus an IF image from human glioma sections (catalog image captions).
Real IHC data
Anti-ELAVL4 Antibody ®
Cat # PB9698
PB9698 is listed for IHC and IF, with human, mouse and rat reactivity (catalog applications and reactivity). Its IHC images show mouse and rat brain and human meningeoma; its IF image shows human glioma (catalog image captions).
Which to pick: Choose PB9698 for tissue IHC on paraffin sections; its mouse brain caption reports citrate retrieval at pH 6 for 20 minutes and primary antibody at 1 μg/ml overnight at 4°C (PB9698 IHC image caption). For IF, PB9698 has a human glioma tissue image; ICC validation is unreported (PB9698 IF image caption; catalog applications). For cross-species IHC, PB9698 has human, mouse and rat images; antibody clonality and section fixative are unreported (PB9698 IHC image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P26378 (ELAV4_HUMAN, ELAV-like protein 4).
- Human Protein Atlas. ELAVL4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ELAVL4 subcellular location (ICC-IF): Localized to the nucleoplasm and vesicles..
- Human Protein Atlas. ELAVL4 antibody validation summary (2 antibodies).
- Mutant FUS and ELAVL4 (HuD) Aberrant Crosstalk in Amyotrophic Lateral Sclerosis. Cell reports 2019 — PMC6613039.
- Isoaspartylation appears to trigger small cell lung cancer-associated autoimmunity against neuronal protein ELAVL4. Journal of neuroimmunology 2016 — PMC5152694.
- Translational derepression of Elavl4 isoforms at their alternative 5' UTRs determines neuronal development. Nature communications 2020 — PMC7125149.
- PubMed PMID:1655278 — UniProt-cited evidence.
- PubMed PMID:12209604 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ELAVL4 (ELAV-like protein 4). Expected localisation Cytoplasmic in brain, adrenal medulla and spermatogonia (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9698). Validated IHC antibody: PB9698 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Medullary cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissues; fixative not specified (datasheet PB9698); 2. Fixation: Image fixative and duration unreported (datasheet PB9698); 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: Citrate pH 6 , 20 min (datasheet PB9698); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9698); 8. Primary antibody: Rabbit anti-ELAVL4, 0.5-1μg/ml (datasheet PB9698); 9. Primary incubation: Overnight at 4 °C (datasheet PB9698). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9698); Caveat: Staining varies by cell type within a section (HPA tissue IHC); Isoform / epitope: Six isoforms; verify antibody epitope coverage (UniProt); Regulation: Brain and adrenal gland enriched RNA (HPA RNA). Evidence: UniProt P26378, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm and vesicles., Datasheet PB9698, PMC6613039, PMC5152694. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ELAVL4/immunohistochemistry-ihc-guide.