EIF4A1 Antibody
Product Specs
Target Background
- Star-PAP-specific polyadenylation sites usage regulates the expression of the eukaryotic translation initiation factor EIF4A1, the tumor suppressor gene PTEN, and the long non-coding RNA NEAT1. PMID: 28911096
- Our findings demonstrate that the shared effects of eIF4A1 and eIF4E on translation are mediated by the coding region and 3'UTR. PMID: 27879264
- Our data identify the eIF4F complex as a critical upstream regulator of TORC1, which acts via TSC2 to inactivate TORC1 upon amino acid withdrawal. PMID: 26988032
- Our results demonstrate that miR-133a plays a pivotal role in colorectal cancer by inhibiting cell proliferation, invasion, and migration by targeting oncogenic eukaryotic translation initiation factor 4A1, which acts as a tumor suppressor and may provide a promising therapeutic target in colorectal cancer. PMID: 28466778
- miR-1284 can function as a novel regulator to reduce gastric cancer multidrug resistant cells by targeting EIF4A1. PMID: 26936591
- Research indicates a growing focus on targeting eukaryotic initiation factor 4A eIF4A1 and eIF4A2 as potential cancer therapies. PMID: 26614665
- Immunohistochemical analysis was performed on over 3000 breast tumors to investigate the relationship between the expression of eIF4A1, the helicase-modulating proteins eIF4B, eIF4E and PDCD4, and clinical outcome. PMID: 25611378
- Collectively, these data suggest a potential role of NS4A in antagonizing host antiviral defense by recruiting eIF4AI and evading the translation inhibition mediated by PKR. PMID: 25866185
- eIF4A functions as an adenosine triphosphate-dependent processive helicase when complexed with two accessory proteins, eIF4G and eIF4B. PMID: 26113725
- Demonstrate that DAP5 associates with eIF2beta and eIF4AI to stimulate IRES-dependent translation of cellular mRNAs. PMID: 25779044
- Studies indicate that eukaryotic initiation factor 4AI (EIF4AI) undergoes significant conformational transitions while unwinding RNA hairpins. PMID: 24909782
- Data indicate that caspase recruitment domain family, member 11 protein CARD11 has complex 5'UTRs and is sensitive to eIF4A RNA helicase inhibition. PMID: 25320244
- Increased Expression of EIF4A1 is associated with a poor response to Brachytherapy in Cervical Cancer. PMID: 24844222
- Report of an eIF4A RNA helicase-dependent mechanism of translational control that contributes to oncogenesis and underlies the anticancer effects of silvestrol and related compounds. PMID: 25079319
- The E186 residue was found to be critical for RNA-dependent ATPase activity for eIF4AI alone and in the presence of eIF4AI-binding domains of eIF4GI. Binding between eIF4AI and eIF4GI domains was also significantly influenced by mutation of E186. PMID: 24471916
- Findings implicate phosphorylation of eIF4G1(Ser1232) by Cdk1:cyclin B and its inhibitory effects on eIF4A helicase activity in the mitotic translation initiation shift. PMID: 24248602
- Eukaryotic translation initiation factor 4A1 (eIF4A1) is a direct target of miR-US25-2-3p. PMID: 23747307
- Regulation of MUC1-C expression is mediated by the PI3K/AKT pathway and the eIF4A RNA helicase. PMID: 22689062
- The results indicated that eIF4AI and eIF4AII expression are linked and that the two protein isoforms exhibit functional differences. PMID: 22589333
- Duplex unwinding and ATPase activities of the DEAD-box helicase eIF4A are coupled by eIF4G and eIF4B. PMID: 21840318
- Study found Burkholderia pseudomallei BPSL1549 acted as a potent cytotoxin against eukaryotic cells; it promotes deamidation of glutamine-339 of the translation initiation factor eIF4A, abolishing its helicase activity and inhibiting translation. PMID: 22076380
- Analysis of the tandem MA-3 region of Pdcd4 protein and characterization of its interactions with eIF4A and eIF4G. PMID: 21454508
- Studies indicate that eIF4A (DDX2), together with its accessory proteins eIF4B and eIF4H, is thought to act as a helicase that unwinds secondary structures in the mRNA 5' UTR. PMID: 21427765
- EIF4A and eIF4G proteins are not responsible for the selective translation of viral mRNAs and the translational shut-off of cellular protein synthesis. PMID: 21377182
- Inhibition of eIF4A reduced the synthesis of APP and tau. PMID: 20927385
- These findings identify PKP1 as a regulator of translation and proliferation via modulation of eIF4A1 activity. PMID: 20156963
- Comparative characterization of two DEAD-box RNA helicases in superfamily II: human translation-initiation factor 4A and hepatitis C virus non-structural protein 3 (NS3) helicase. PMID: 11903057
- Results indicate that not only binding to eIF4A but also prevention of eIF4A binding to the MA-3 domain of eIF4Gc contributes to the mechanism by which Pdcd4 inhibits translation. PMID: 15082783
- Analysis of cells of eIF4GIf molecules lacking either the PABP-binding site, the eIF3-binding site, the middle domain eIF4A-binding site, or the C-terminal segment that includes the second eIF4A-binding site. PMID: 17130132
- Results report that 15d-PGJ2 blocks translation through inactivation of translational initiation factor eIF4A. PMID: 18034160
- The PDCD4 MA3 domains compete with the eIF4G MA3 domain and RNA for eIF4A binding. PDCD4 inhibits translation initiation by displacing eIF4G and RNA from eIF4A. PMID: 18296639
- The interaction of eIF4AI with two accessory factors, eIF4B and eIF4H, was studied. PMID: 18719248
- Study reports the topology of the eIF4A/4G/4H helicase complex, which is built from multiple experimentally observed domain-domain contacts. PMID: 19203580
Q&A
What experimental strategies ensure specificity when selecting EIF4A1 antibodies for Western blotting?
EIF4A1 antibody specificity must be validated through orthogonal methods due to its high homology with EIF4A2 (79% amino acid identity). Key steps include:
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Immunogen alignment: Select antibodies targeting non-conserved regions (e.g., residues 250-407 in human EIF4A1) .
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Knockout validation: Use Eif4a1 / Eif4a2 double-knockout cell lysates as negative controls .
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Cross-reactivity profiling: Test against recombinant EIF4A2 and other DEAD-box helicases (e.g., DDX3X, DDX5).
Table 1: Performance metrics of common EIF4A1 antibody clones
| Clone | Epitope Region | Cross-Reactivity | Applications Validated |
|---|---|---|---|
| EPR14506 | C-terminal | EIF4A2 (weak) | WB, IP, IF |
| 7C3 | ATP-binding | None reported | ELISA, IHC |
| YA774 | N-terminal | Mouse ortholog | WB, functional assays |
How does EIF4A1's role in translation initiation impact experimental design in B cell studies?
EIF4A1 facilitates ribosome scanning through structured 5'UTRs of mRNAs encoding cell cycle regulators (e.g., MYC, CCND1). In B cell activation studies:
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Timing considerations: Protein synthesis rates increase 2.4-fold within 6 hours of BCR stimulation .
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Dosage controls: Use heterozygous Eif4a1 + / − cells to avoid compensatory EIF4A2 upregulation .
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Functional readouts: Combine polysome profiling with siRNA knockdown to identify EIF4A1-dependent transcripts.
What validation protocols are essential for immunohistochemistry (IHC) applications?
Three-tier validation is critical:
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Genetic confirmation: Compare staining in wild-type vs. Eif4a1 − / − tissues .
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Subcellular localization: Verify nuclear-cytoplasmic distribution matches IF data .
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Pre-absorption controls: Demonstrate ≥90% signal reduction with immunogen peptides .
How to resolve contradictory data on EIF4A1's essentiality in different cell types?
The PMC study reveals context-dependent requirements :
Table 2: Tissue-specific EIF4A1 dependency
| Cell Type | Viability Post-KO | Compensatory Mechanism |
|---|---|---|
| Germinal Center B cells | Non-viable | None detected |
| Hepatocytes | Viable | EIF4A2 upregulation (4.7-fold) |
| Fibroblasts | Partially viable | EIF4A3 recruitment |
Methodological recommendations:
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Perform concurrent EIF4A1/EIF4A2 quantification via mass spectrometry
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Use inducible CRISPR systems to avoid developmental compensation
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Validate with small-molecule inhibitors (Hippuristanol > Rocaglamide A specificity)
What advanced techniques elucidate EIF4A1's structural interactions?
Cryo-EM-guided mutagenesis:
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Map antibody-epitope interfaces using 3.2Å resolution structures
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Design point mutations (e.g., K238A) disrupting ATPase activity
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Perform in vitro translation assays with structured GFP reporters
Single-molecule FRET:
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Quantify helicase unwinding rates (∼35 bp/s under physiological ATP)
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Test antibody effects on conformational changes during RNA binding
How to analyze isoform-specific functions in CRISPR-edited models?
Dual knockout strategy:
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Generate Eif4a1 − / −Eif4a2 + / − hypomorphic cells
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Introduce siRNA-resistant cDNA variants
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Monitor translation fidelity via Ribo-seq
Critical parameters:
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Maintain [ATP]/[ADP] ratios at 3:1 to prevent artifactual helicase stalling
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Use TIDE analysis to confirm editing efficiency (>95% recommended)
Quantitative framework for translational control studies
Application guidelines:
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Normalize to actinomycin D-treated controls
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Use cycloheximide (100μg/mL, 10min) to freeze ribosome positions
Multi-parametric antibody validation matrix
Scoring criteria:
| Parameter | Weight | Assessment Method |
|---|---|---|
| Epitope uniqueness | 30% | BLAST vs. human proteome |
| Batch consistency | 25% | CV <15% across 3 lots |
| Functional impact | 20% | Helicase activity inhibition |
| Cross-species | 15% | Primate vs. rodent reactivity |
| Long-term stability | 10% | 12-month accelerated aging |
