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ELAVL4 / ELAV-like protein 4 · IHC design guide
Design Immunohistochemistry for ELAVL4
Plan chromogenic ELAVL4 IHC in paraffin sections using the catalog antibody’s documented conditions. Compare cytoplasmic staining with the reported tissue pattern and assess cell identity when scoring (HPA tissue IHC; datasheet PB9698).
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 | Cytoplasm (HPA tissue IHC); dendrites and axons (UniProt) | |
| Staining pattern | Cytoplasmic staining in neurons, adrenal medulla 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 | Spermatogonia also stain; assess cell identity (HPA tissue IHC) | |
| Regulation | No stimulus-linked expression change is established (UniProt) | |
| Isoform / epitope | 6 isoforms; epitope coverage needs validation (UniProt) |
Section 1
Recommended ELAVL4 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PB9698) is followed by published ELAVL4 staining methods for human spinal cord (PMC6613039) and small cell lung cancer sections (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 retrieval in citrate buffer at pH 6 (datasheet: PB9698); the spinal cord protocol specifies 120°C for 10 minutes (PMC6613039).
Section 2
What Is the Expected ELAVL4 Staining Pattern?
ELAVL4 should show cytoplasmic staining in neuronal cells of the brain and medullary cells of the adrenal gland, with staining also reported in testicular spermatogonia (HPA: tissue IHC, Enhanced reliability). Signal may extend through the perikaryon and neuronal projections (UniProt P26378: subcellular location). ELAVL4 has no transmembrane segment, so a crisp cell-surface rim is not the expected pattern (UniProt P26378: topology).
| Cytoplasmic staining in neurons of the cortex, caudate or hippocampus, with visible cell bodies and processes. | This fits the reported high neuronal staining in these regions (HPA: tissue IHC) and the perikaryon, dendrite and axon locations (UniProt P26378: subcellular location). Judge the labelled cells and compartment together; a brown area alone does not identify the cell type. |
| Predominantly nuclear staining, or a sharp outline confined to the plasma membrane, in a paraffin section. | Neither matches the reported cytoplasmic tissue pattern (HPA: tissue IHC); ELAVL4 has no transmembrane segment (UniProt P26378: topology). Check morphology and controls before calling it target signal. Nuclear IF images are a separate observation (HPA: subcellular ICC-IF). |
| Strong staining in adipocytes, respiratory epithelium or hematopoietic cells while expected positive cells are absent. | Those cell types are listed as not detected (HPA: tissue IHC). The pattern raises concern for antibody cross-reactivity or endogenous detection activity (general IHC practice). Compare a matched negative control and inspect whether the signal tracks a cell type or the detection reaction. |
| Diffuse color covers several structures without a clear neuronal, medullary-cell or spermatogonial pattern. | This does not reproduce the reported cell-restricted cytoplasmic pattern (HPA: tissue IHC). Diffuse background can arise from nonspecific antibody binding or detection chemistry (general IHC practice); it should not be scored as ELAVL4-positive cells. |
| No convincing staining in a section containing identifiable brain neurons or adrenal medullary cells. | These cells are reported as high staining (HPA: tissue IHC). Verify that they are present, then review the catalog antibody’s IHC-P instructions and run controls for the detection workflow (general IHC practice). A blank section alone cannot distinguish absent target from assay failure. |
💡Expected ELAVL4 appearanceCall a positive result when cytoplasmic signal is clearly stronger in brain neurons or adrenal medullary cells than in surrounding cells (HPA: tissue IHC); diffuse color or an isolated membrane rim is suspect (HPA: tissue IHC; UniProt P26378: topology).
| Tissue and cell selection | High staining is reported in caudate, cerebral cortex and hippocampal neurons, adrenal medullary cells, and testicular spermatogonia (HPA: tissue IHC). Select and score the named cells; neighboring cells need not share their staining. |
| IHC antibody evidence | The tissue profile has Enhanced reliability, meaning high consistency between staining and RNA expression data (HPA: tissue IHC). CAB004442 has Enhanced IHC validation; HPA043047 is listed as Approved for ICC, with no IHC status supplied (HPA: antibodies). Validation for one assay does not establish the other. |
| Intracellular distribution | UniProt places ELAVL4 in cytoplasm, perikaryon, dendrites, axons and growth cones; it also reports association with polysomes (UniProt P26378: subcellular location). Assess cytoplasm and neuronal processes where preserved, without requiring every thin projection to be visible in a section. |
| IF/ICC Q: Is its pattern identical to tissue IHC? | A: HPA reports approved nucleoplasm and vesicle localization in ICC-IF images, while its tissue IHC profile reports cytoplasmic staining (HPA: subcellular ICC-IF; HPA: tissue IHC). Record the assay and compartment when comparing results; the ICC-IF observation does not redefine the expected paraffin-section pattern. |
| Isoforms and antibody epitope | UniProt lists six ELAVL4 isoforms and three RNA recognition motifs (UniProt P26378: isoforms, domains). The supplied records do not locate either antibody’s epitope, so they do not support a prediction that one isoform or domain will stain more strongly. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control brain or adrenal tissue is blank. | The expected high-staining cells may be missing from the field (HPA: tissue IHC), or an IHC workflow step may have failed (general IHC practice). | Locate identifiable neurons or medullary cells first (HPA: tissue IHC). Then check the catalog antibody’s IHC-P instructions, detection reagents and a concurrent assay control (general IHC practice). No ELAVL4-specific fixation sensitivity is supplied. |
| Signal is nuclear only in tissue IHC. | This differs from the cytoplasmic tissue profile (HPA: tissue IHC); nuclear localization is reported separately for ICC-IF (HPA: subcellular ICC-IF). | Confirm the section’s cell morphology and compare appropriate detection controls (general IHC practice). Report the observed compartment and assay; do not use the ICC-IF result alone to validate nuclear tissue staining. |
| Many unrelated cell types stain uniformly. | The distribution exceeds the named positive cells (HPA: tissue IHC) and may reflect nonspecific binding or endogenous detection activity (general IHC practice). | Compare a matched control lacking primary antibody, review blocking and detection steps, and rescore only identifiable cells with compartment-specific signal (general IHC practice). |
| A negative-reference cell population stains strongly. | Adipocytes, bronchial respiratory epithelium and bone-marrow hematopoietic cells are listed as not detected (HPA: tissue IHC). | Check those cells against a known-positive section in the same run (HPA: tissue IHC; general IHC practice). If the unexpected pattern persists, assess antibody specificity and detection controls before interpreting it as ELAVL4. |
| Neuronal processes are faint although cell bodies stain. | ELAVL4 is reported in perikaryon and neuronal projections (UniProt P26378: subcellular location), but thin processes can be hard to resolve in sections (general IHC practice). | Inspect well-preserved areas at suitable magnification and score identifiable cell bodies separately from visible processes (general IHC practice). Do not require every axon or dendrite to stain in a single section. |
| An IF/ICC image appears inconsistent with the IHC slide. | HPA reports nucleoplasm and vesicles for ICC-IF but cytoplasm for tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). | Compare results within their own assay and antibody validation status (HPA: antibodies). Document the cell type and compartment seen; use the separate IF/ICC guide for that assay’s workflow. |
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; neuronal cells should stain (HPA: cerebral cortex neuronal cells High). Use adipose tissue as the negative, where adipocytes are not detected (HPA: adipose tissue adipocytes Not detected); on the cortex slide, use morphologically non-neuronal cells as provisional internal negatives that should show only background staining (HPA: cerebral cortex neuronal cells High; standard IHC practice).
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 control, a nonimmune rabbit IgG control matched to the primary antibody’s clonality, and ELAVL4-knockout tissue or a validated peptide-block control (caption: rabbit anti-ELAVL4 antibody; standard IHC practice). For DAB detection in brain, block endogenous peroxidase and assess endogenous biotin when using the caption’s biotin-based detection system (caption: biotinylated secondary and SABC with DAB; standard IHC practice).
⚠️Feasibility: The PB9698 mouse-brain paraffin-section caption reports 20 minutes of heat retrieval in citrate buffer at pH 6, but does not establish retrieval dependence or report a fixative; a target-specific fixation window is unreported (PB9698 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; the HPA ICC-IF images document cultured cell lines, not tissue IF (HPA: subcellular). In brain sections, neuronal lipofuscin can complicate interpretation of brown chromogen and can autofluoresce in IF, so check pigment and background controls (standard IHC/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, compartment, controls and scoring against the available tissue evidence.
What retrieval condition should I start with for ELAVL4 in paraffin sections?
Use heat-mediated citrate pH 6 antigen retrieval for 20 minutes before staining paraffin sections (datasheet PB9698; PB9698 tissue-IHC caption). The demonstrated mouse brain workflow used 10% goat serum, 1 µg/mL catalog antibody overnight at 4 °C, then biotinylated secondary and DAB (PB9698 tissue-IHC caption). If staining is weak, compare a small, controlled increase in retrieval time while watching for tissue damage and diffuse background (standard IHC practice). Keep section thickness, antibody concentration and detection conditions matched across the comparison so a retrieval change remains interpretable (standard IHC practice).
Could fixation explain weak or uneven ELAVL4 staining?
The selected tissue caption identifies paraffin-embedded mouse brain but does not state its fixative, so target-specific fixation sensitivity is unknown (PB9698 tissue-IHC caption). Neither the tissue staining profile nor the protein’s topology and modifications establish how a particular fixative affects its epitope (HPA tissue IHC; UniProt P26378). For a controlled comparison, record fixation method and duration, section thickness and storage, then stain matched sections together using citrate pH 6 retrieval (standard IHC practice; datasheet PB9698). Inspect poorly preserved areas separately from well-preserved cells, and compare positive controls before changing antibody concentration (standard IHC practice).
Which staining compartments support an ELAVL4 call?
In tissue IHC, look first for cytoplasmic staining in appropriate cells: the reported profile includes brain, adrenal medulla and spermatogonia (HPA tissue IHC). ELAVL4 is also annotated in perikarya, dendrites, axons and growth cones, consistent with staining that extends beyond the cell body (UniProt P26378 subcellular location). Approved ICC/IF locations include nucleoplasm and vesicles, so nuclear signal alone needs assay-specific controls rather than automatic rejection (HPA subcellular). Compare each compartment with adjacent cells, a no-primary control and tissue morphology; avoid calling diffuse staining across damaged or acellular areas positive (standard IHC practice).
How can epitope choice affect ELAVL4 IHC results?
ELAVL4 has 6 annotated isoforms and three RNA recognition motifs at residues 51–129, 137–217 and 302–380 (UniProt P26378). Without an epitope map for the catalog antibody, an absent stain cannot establish that every isoform is absent, and an apparently restricted pattern cannot identify an isoform (UniProt P26378; standard IHC interpretation). Phosphorylation and alternative methylation are annotated, but their effects on this antibody’s staining have not been supplied (UniProt P26378 modified residues). If epitope coverage matters, obtain the antibody’s immunogen information and compare it with isoform sequences before interpreting differences between specimens (standard IHC practice).
How should I adapt the ELAVL4 question to multiplex IF?
Use a marker for the expected cell population alongside ELAVL4, such as an independently validated neuronal marker in brain, to establish which cells carry the signal (HPA tissue IHC; standard IF practice). Choose fluorophores after checking unstained tissue autofluorescence, and place the weaker signal in a cleaner spectral channel (standard IF practice). ELAVL4 has no transmembrane segment and is annotated in cytoplasm and cell projections; use controlled permeabilisation to reach intracellular epitopes (UniProt P26378 topology and subcellular location; standard IF practice). Optimise IF fixation and permeabilisation with IF controls because the PB9698 tissue caption reports a chromogenic paraffin-section workflow, not IF conditions (PB9698 tissue-IHC caption).
What should I check when DAB background obscures ELAVL4?
Run a no-primary control through the same secondary and detection steps to locate reagent-derived background (standard IHC practice). The documented workflow uses a biotinylated goat anti-rabbit secondary, a streptavidin-biotin complex and DAB; check biotin-related signal and block endogenous peroxidase as appropriate for that detection system (PB9698 tissue-IHC caption; standard IHC practice). Inspect tissue edges, folds and damaged regions separately, since localized artefact can mimic cellular staining (standard IHC practice). If background remains widespread, titrate the primary below the demonstrated 1 µg/mL and compare matched sections while holding retrieval and DAB development constant (PB9698 tissue-IHC caption; standard IHC practice).
How should I score ELAVL4 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because ELAVL4 tissue staining is reported in brain, adrenal medulla and spermatogonia, while subcellular annotations span cytoplasm and projections (HPA tissue IHC; UniProt P26378). For cellular staining, report the percentage of positive target cells or an H-score using fixed intensity bins; for a prespecified projection-rich area, consider positive area per mm² (standard IHC practice). Normalize cell-based scores to eligible cells counted and area-based scores to viable tissue area, excluding folds and necrosis (standard IHC practice). Apply one threshold, counterstain and image-acquisition setup across batches, with control sections to track staining drift (standard IHC practice).
How can I distinguish true ELAVL4 staining from artefact?
A convincing tissue result combines cellular morphology with the reported cytoplasmic pattern in brain, adrenal medulla or spermatogonia (HPA tissue IHC). Signal in perikarya or neuronal projections can also fit the annotated distribution, while the ICC/IF record includes nucleoplasm and vesicles and warrants compartment-specific validation (UniProt P26378 subcellular location; HPA subcellular). Treat uniform staining at section edges, in necrotic areas or in the no-primary control as suspect, and assess endogenous peroxidase contribution to DAB signal (standard IHC practice). Compare positive and low-expression reference cells on the same run before assigning biological meaning to an isolated stain (HPA tissue IHC; standard IHC practice).
Boster reagents
Best ELAVL4 / ELAV-like protein 4 IHC Antibodies
PB9698 has IHC data from paraffin sections of mouse and rat brain and human meningeoma, plus IF data from paraffin sections of human glioma (catalog image captions).
Real IHC data
Anti-ELAVL4 Antibody ®
Cat # PB9698
PB9698 is listed for IHC and IF in human, mouse and rat (catalog applications and reactivity). Its IHC captions show mouse brain, rat brain and human meningeoma; its IF caption shows human glioma (catalog image captions).
Which to pick: For tissue IHC, choose PB9698: its own IHC captions document paraffin sections, and the mouse brain example used 1 μg/ml after citrate retrieval (PB9698 IHC captions). For IF, PB9698 has a human glioma section image; ICC validation is unreported (PB9698 IF caption and applications). PB9698 lists human, mouse and rat reactivity, while clonality and the tissue fixative are unreported (catalog reactivity, clone field and image captions).
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 Cytoplasm (HPA tissue IHC); dendrites and axons (UniProt). 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: Spermatogonia also stain; assess cell identity (HPA tissue IHC); Isoform / epitope: 6 isoforms; epitope coverage needs validation (UniProt); Regulation: No stimulus-linked expression change is established (UniProt). 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.