This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
EIF4A2 / Eukaryotic initiation factor 4A-II · IHC design guide
Design Immunohistochemistry for EIF4A2
Plan EIF4A2 chromogenic IHC in paraffin sections using the cytoplasmic tissue pattern as a starting point (HPA tissue IHC). The catalog antibody’s IHC range is 0.5–1 μg/mL (datasheet PB9758); interpret staining with controls because tissue evidence has uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EIF4A2 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 | Ubiquitous cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across diverse tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9758) | |
| Positive control | Adipose tissue+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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9758) | |
| Caveat | Staining reliability is uncertain; validate the pattern (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended EIF4A2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: PB9758) with published EIF4A2 IHC methods (PMC7471732; PMC6518650; PMC13366719).
| Sample | Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet PB9758) |
| Fixation | Image fixative and duration unreported (datasheet PB9758); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet PB9758); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9758) |
| Primary antibody | Rabbit anti-EIF4A2, 0.5-1μg/ml (datasheet PB9758) |
| Primary incubation | Overnight at 4 °C (datasheet PB9758) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9758) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EIF4A2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: PB9758); optimize staining with positive and negative controls (standard IHC practice).
Section 2
What Is the Expected EIF4A2 Staining Pattern?
EIF4A2 should appear mainly in the cytoplasm of many cell types in paraffin sections (HPA: ubiquitous cytoplasmic expression). Reported high staining includes adipocytes, glandular cells, respiratory epithelial cells and selected brain cells (HPA: tissue IHC). The protein has no transmembrane segment (UniProt Q14240 topology). Treat the pattern as provisional: HPA rates tissue IHC reliability Uncertain, with external verification pending (HPA: reliability).
| Cytoplasmic staining in adipocytes, glandular cells or respiratory epithelial cells. | This fits reported high staining in adipose tissue, adrenal gland, appendix, breast and bronchus (HPA: tissue IHC). Compare cells within the section; high is an HPA category, not a universal chromogen intensity threshold (HPA: tissue IHC; standard IHC practice). |
| Strong nuclear-only or membrane-rim staining with little cytoplasmic signal. | This conflicts with the reported cytoplasmic tissue pattern and approved cytosol IF location (HPA: tissue IHC; HPA: subcellular). Check morphology and controls before calling it EIF4A2; nonspecific staining or detection artefact is possible (standard IHC practice). |
| Signal confined to an unexpected cell population while expected cells remain unstained. | Consider cross-reactivity or endogenous detection activity (standard IHC practice). Unlisted cells are not established negatives: HPA describes ubiquitous expression, and its tissue IHC reliability is Uncertain (HPA: tissue IHC; HPA: reliability). |
| Diffuse color over cells and surrounding tissue, obscuring cell boundaries. | A uniform haze cannot establish cytoplasmic localisation (standard IHC practice). Background from detection reagents, incomplete blocking or excess primary antibody is possible; compare a matched negative control before scoring cells (standard IHC practice). |
| No signal in adipocytes or a listed high-staining glandular cell population. | This disagrees with the reported pattern (HPA: High in adipocytes and glandular cells). Review the section and assay controls before concluding biological absence; HPA's Uncertain reliability also limits any single reference tissue as proof (HPA: reliability; standard IHC practice). |
💡Expected EIF4A2 appearanceCall a result consistent when interpretable cytoplasmic staining is present in reported high-staining cells; isolated nuclear, membrane-rim or tissue-wide haze is suspect (HPA: tissue IHC; HPA: subcellular; standard IHC practice).
| Reference cell populations | HPA reports High staining in adipocytes, several glandular epithelia, bronchial respiratory epithelium, glia and cerebellar granular-layer cells (HPA: tissue IHC). Use these as comparison populations, not as guaranteed controls, because tissue IHC reliability is Uncertain (HPA: reliability). |
| Compartment and topology | The tissue pattern is cytoplasmic and ICC-IF places EIF4A2 in the cytosol (HPA: tissue IHC; HPA: subcellular). UniProt lists no transmembrane segment, signal peptide or propeptide (UniProt Q14240 topology/processing); these facts do not predict staining intensity. |
| Antibody validation | The listed IHC antibody CAB011690 is rated Uncertain, while HPA068286 is Approved for ICC rather than IHC (HPA: antibodies). Agreement between assay types is useful context, but their validation labels apply to different antibodies and applications (HPA: antibodies). |
| Isoforms and epitope coverage | UniProt lists two EIF4A2 isoforms and helicase regions at residues 64–235 and 246–407 (UniProt Q14240 isoforms/domains). Without an epitope map for the IHC antibody, neither isoform coverage nor a domain-specific staining difference can be assigned. |
| IF/ICC cross-check? | Yes: HPA reports approved cytosol localisation in ICC-IF, with images from CACO-2, U-251MG and U2OS (HPA: subcellular). Its summary cautions that the localisation uses antibodies targeting proteins from multiple genes; it does not validate a new IHC result (HPA: subcellular; HPA: antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed high-staining cell population appears blank. | Possible assay failure or weak detection; the HPA High category is an observed reference, with Uncertain IHC reliability (HPA: tissue IHC; HPA: reliability; standard IHC practice). | Confirm cell identity, inspect a concurrent positive control, then review retrieval, primary antibody incubation and chromogen development against the chosen assay instructions (standard IHC practice). |
| Predominantly nuclear staining is scored as positive. | The compartment disagrees with cytoplasmic tissue staining and approved cytosol ICC-IF localisation (HPA: tissue IHC; HPA: subcellular). | Recheck cell boundaries with the counterstain and examine negative controls; report the discordance rather than counting nuclear-only cells as confirmed positive (standard IHC practice). |
| Membrane outlines dominate the section. | A membrane-rim pattern is unsupported by the reported localisation and the absence of a transmembrane segment (HPA: tissue IHC; UniProt Q14240 topology). | Inspect morphology and controls for edge or detection artefact; seek an independently validated IHC reagent if the pattern persists (standard IHC practice; HPA: antibodies). |
| Color appears in many cell types but lacks clear intracellular detail. | Broad distribution may be real, but diffuse background can hide localisation (HPA: ubiquitous cytoplasmic expression; standard IHC practice). | Compare a matched negative control, review blocking and washing, and adjust primary antibody concentration using the assay instructions before scoring (standard IHC practice). |
| An unexpected cell type stains strongly while expected cells do not. | Cross-reactivity or endogenous detection activity is possible; HPA supplies no negative tissue list and rates IHC reliability Uncertain (HPA: tissue IHC; HPA: reliability; standard IHC practice). | Check a no-primary control for detection activity and assess a second validated antibody or orthogonal evidence before assigning cell-specific expression (standard IHC practice). |
| ICC-IF looks cytosolic, but the paraffin IHC pattern differs. | HPA's Approved ICC and Uncertain IHC labels belong to different listed antibodies and assay settings (HPA: antibodies; HPA: subcellular). | Interpret each assay against its own controls and validation status; use the IF result as a localisation cross-check, not as proof that the IHC signal is specific (standard IHC practice; HPA: antibodies). |
Experiment evidence
IHC & IF images showing EIF4A2 staining
PB9758 · IHCIHC analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758). eIF4A2 was detected in a paraffin-embedded section of mouse spleen tissue. Heat
PB9758 · IHCIHC analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758). eIF4A2 was detected in a paraffin-embedded section of rat intestine tissue. Heat
PB9758 · IFIF analysis of eIF4A2 using anti-eIF4A2 antibody (PB9758). eIF4A2 was detected in immunocytochemical section of A431 cell. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156976-EIF4A2/tissue/cerebral+cortex#imid_3704064
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156976-EIF4A2/tissue/colon#imid_3703990
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156976-EIF4A2/tissue/liver#imid_3704007
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156976-EIF4A2/tissue/kidney#imid_3704000
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA068286; green = EIF4A2, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA068286; green = EIF4A2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EIF4A2 IHC & IF
🧪Run bronchus first and look for staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells). HPA detects EIF4A2 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and do not designate any cell type on the positive slide as internally negative without independent validation (HPA: no negative tissues).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: EIF4A2 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 EIF4A2 in CACO-2, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Use secondary-only and host- and clonality-matched rabbit IgG isotype controls, plus EIF4A2 knockout tissue if available, to assess specificity (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and check for background from bronchial inflammatory cells before interpreting DAB staining (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected paraffin-section caption does not state a fixative (caption: fixative unreported). Heat retrieval with EDTA at pH 8.0 was used for mouse spleen, but its necessity in bronchus is unreported (caption: EDTA retrieval in mouse spleen). The supplied evidence does not establish that frozen sections or IF/ICC are easier; HPA reports cytosolic ICC-IF staining with a multiple-gene antibody caution, while bronchial mucus can contribute background in chromogenic IHC (HPA: cytosol, multiple-gene caution; standard IHC practice).
HPA tissue IHC evidence for EIF4A2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: EIF4A2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced EIF4A2 IHC Tips
Troubleshoot EIF4A2 staining in paraffin sections by checking retrieval, compartment, controls and cell-level scoring before interpreting chromogenic signal.
What retrieval should I try first if EIF4A2 staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet PB9758). That condition was used for the catalog antibody’s mouse spleen image, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet PB9758). If staining remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Check tissue preservation and the positive control before increasing retrieval intensity, because excessive heating can damage morphology and make cytoplasmic staining harder to judge (standard IHC practice). Record both signal and tissue integrity for each condition (standard IHC practice).
Could fixation be masking EIF4A2 in my paraffin sections?
EIF4A2-specific sensitivity to fixation is unknown from the supplied evidence; the catalog image identifies a paraffin section but does not state its fixative (datasheet PB9758). Document the specimen’s actual fixative and processing history before comparing staining across samples (standard IHC practice). Use similarly processed control sections when assessing a weak result, because fixation and processing differences can alter antigen accessibility in IHC (standard IHC practice). Apply the stated EDTA pH 8.0 heat retrieval consistently while investigating those differences (datasheet PB9758). Do not attribute a change in signal to EIF4A2 fixation sensitivity without a controlled comparison (standard IHC practice).
Where should EIF4A2 staining appear, and how should I assess nuclear signal?
Prioritise a cytoplasmic staining pattern when reviewing paraffin sections, since HPA reports ubiquitous cytoplasmic tissue expression and approved cytosol localisation (HPA tissue IHC; HPA subcellular). UniProt does not annotate a subcellular location for EIF4A2, so the HPA pattern should guide review rather than serve as independent UniProt confirmation (UniProt Q14240; HPA subcellular). Compare nuclear and cytoplasmic signal in intact cells on the same section, using the counterstain to define cell boundaries (standard IHC practice). If nuclear staining dominates, check antibody omission and detection controls, then repeat the comparison on well-preserved tissue (standard IHC practice). HPA flags its subcellular assignment as based on antibodies targeting proteins from multiple genes (HPA subcellular).
How can I tell whether the antibody detects both EIF4A2 isoforms?
UniProt lists 2 EIF4A2 isoforms, but the supplied antibody caption does not define the recognised epitope or establish isoform coverage (UniProt Q14240; datasheet PB9758). Request the immunogen sequence or epitope map and compare it with each isoform before claiming isoform-specific staining (standard antibody validation practice; UniProt Q14240). EIF4A2 has no annotated transmembrane segment, and its recorded helicase domains span residues 64–235 and 246–407 (UniProt Q14240). Phosphothreonine at residue 159 is annotated, but its effect on this antibody’s binding has not been established by the supplied evidence (UniProt Q14240; datasheet PB9758). Use a validated specificity control to investigate unexpected staining (standard IHC practice).
How should I compare multiplex IF with this chromogenic EIF4A2 result?
Treat multiplex IF as a separate assay when comparing it with the paraffin-section IHC result; the supplied catalog image documents chromogenic detection with DAB (datasheet PB9758). Pair EIF4A2 with a validated marker for the expected cell type, and compare cellular overlap rather than assuming that nearby signals come from the same cell (standard IF practice). Choose fluorophores and imaging channels after checking tissue autofluorescence with an unstained control (standard IF practice). For an intracellular epitope, optimise permeabilisation in the IF assay; EIF4A2 has no annotated transmembrane segment, while HPA reports cytosol localisation (standard IF practice; UniProt Q14240; HPA subcellular). Keep IF exposure and IHC scoring decisions documented separately (standard imaging practice).
What should I check when the whole section looks brown?
First compare the stained section with an antibody-omission control to identify signal from detection chemistry or tissue rather than the primary antibody (standard IHC practice). The documented workflow uses 10% goat serum blocking, a biotinylated secondary antibody, a streptavidin–biotin complex and DAB (datasheet PB9758). Check peroxidase blocking as a general DAB workflow step, and consider a detection control for endogenous biotin when using that documented amplification method (standard IHC practice; datasheet PB9758). Review section edges, damaged areas and precipitate separately from intact cell cytoplasm (standard IHC practice). Lower primary concentration only after controls identify antibody-dependent background, keeping retrieval and development conditions matched (standard IHC practice).
How should I score EIF4A2 across paraffin-section samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring rule before reviewing groups, since HPA describes a ubiquitous cytoplasmic tissue pattern (standard IHC practice; HPA tissue IHC). An H-score combines the percentage of cells at each staining intensity; report the percentage of positive cells alongside it when possible (standard IHC practice). If counting stained cells, express density per mm² of viable analysed tissue and exclude artefactual or necrotic areas by a predefined rule (standard IHC practice). Normalise comparisons to the same cell type or annotated tissue area, with matched retrieval, detection and image acquisition conditions (standard IHC practice). Report scoring thresholds and observer agreement so differences can be assessed (standard IHC practice).
How do I distinguish genuine EIF4A2 signal from tissue artefact?
Look for staining in intact cell cytoplasm, consistent with HPA’s tissue expression profile and approved cytosol localisation (HPA tissue IHC; HPA subcellular). Do not assume a positive cell type is exclusive to EIF4A2: HPA reports low tissue specificity and marks its tissue IHC reliability as uncertain (HPA tissue IHC). Compare suspicious edge or necrotic staining with preserved regions, and use an antibody-omission control to assess endogenous enzyme or detection signal (standard IHC practice). The catalog mouse spleen image demonstrates staining after EDTA pH 8.0 retrieval and DAB development, but does not establish specificity for every specimen (datasheet PB9758). Resolve discordant patterns with an independently validated specificity control (standard IHC practice).
Boster reagents
Best EIF4A2 / Eukaryotic initiation factor 4A-II IHC Antibodies
PB9758 has IHC data from paraffin sections of mouse spleen, rat intestine, and human mammary cancer (IHC image captions), plus IF data from A431 cells (IF image caption).
Real IHC data
Anti-eIF4A2 Antibody ®
Cat # PB9758
PB9758 is listed for IHC in human, mouse, and rat, with paraffin-section images from all three species (catalog applications and reactivity; IHC image captions). PB9758 is also listed for IF and ICC, with an IF image from A431 cells (catalog applications; IF image caption).
Which to pick: For tissue IHC, choose PB9758: its mouse spleen paraffin-section image documents EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody (PB9758 IHC image caption); the fixative is unreported (PB9758 IHC image caption). For IF/ICC, PB9758 has an A431-cell IF example at 2 μg/ml (PB9758 IF image caption). For cross-species IHC, PB9758 is listed as reactive with human, mouse, and rat, with paraffin-section images for each (catalog reactivity; PB9758 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14240 (IF4A2_HUMAN, Eukaryotic initiation factor 4A-II).
- Human Protein Atlas. EIF4A2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. EIF4A2 subcellular location (ICC-IF): Localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. EIF4A2 antibody validation summary (2 antibodies).
- High expression of eIF4A2 is associated with a poor prognosis in esophageal squamous cell carcinoma. Oncology letters 2020 — PMC7471732.
- Eukaryotic initiation factor 4A2 promotes experimental metastasis and oxaliplatin resistance in colorectal cancer. Journal of experimental & clinical cancer research : CR 2019 — PMC6518650.
- Long non-coding RNA LINC00520 promotes the proliferation and metastasis of malignant melanoma by inducing the miR-125b-5p/EIF5A2 axis. Journal of experimental & clinical cancer research : CR 2020 — PMC7254730.
- Increased mRNA translation delays tumour initiation and exposes a therapeutic vulnerability in lung cancer. Molecular cancer 2026 — PMC13366719.
- PubMed PMID:8521730 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EIF4A2 (Eukaryotic initiation factor 4A-II). Expected localisation Ubiquitous cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9758). Validated IHC antibody: PB9758 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet PB9758); 2. Fixation: Image fixative and duration unreported (datasheet PB9758); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet PB9758); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9758); 8. Primary antibody: Rabbit anti-EIF4A2, 0.5-1μg/ml (datasheet PB9758); 9. Primary incubation: Overnight at 4 °C (datasheet PB9758). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9758); Caveat: Staining reliability is uncertain; validate the pattern (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q14240, HPA tissue IHC · Uncertain, HPA ICC-IF · cytosol. Caution: Based on antibodies targeting proteins from…, Datasheet PB9758, PMC7471732, PMC6518650. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EIF4A2/immunohistochemistry-ihc-guide.