Anti-eIF4A3 antibody (ab32485)
- Product nameAnti-eIF4A3 antibodySee all eIF4A3 primary antibodies ...
- DescriptionRabbit polyclonal to eIF4A3
- Tested applicationsIHC-P, IP, ICC/IF, WB more details
- Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Xenopus laevis, Zebrafish, Cynomolgus Monkey
Synthetic peptide derived from within residues 1 - 100 of Human eIF4A3.
(Peptide available as ab32484.)
- Positive controlThis antibody gave a positive signal in the following lysates: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab32485 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||IP: Use a concentration of 5 µg/ml.|
|ICC/IF||ICC/IF: Use a concentration of 5 µg/ml.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).Can be blocked with eIF4A3 peptide (ab32484).|
- FunctionATP-dependent RNA helicase. Component of a splicing-dependent multiprotein exon junction complex (EJC) deposited at splice junction on mRNAs. The EJC is a dynamic structure consisting of a few core proteins and several more peripheral nuclear and cytoplasmic associated factors that join the complex only transiently either during EJC assembly or during subsequent mRNA metabolism. Core components of the EJC, that remains bound to spliced mRNAs throughout all stages of mRNA metabolism, functions to mark the position of the exon-exon junction in the mature mRNA and thereby influences downstream processes of gene expression including mRNA splicing, nuclear mRNA export, subcellular mRNA localization, translation efficiency and nonsense-mediated mRNA decay (NMD). Constitutes at least part of the platform anchoring other EJC proteins to spliced mRNAs. Its RNA-dependent ATPase and RNA-helicase activities are induced by CASC3, but abolished in presence of the MAGOH/RBM8A heterodimer, thereby trapping the ATP-bound EJC core onto spliced mRNA in a stable conformation. The inhibition of ATPase activity by the MAGOH/RBM8A heterodimer increases the RNA-binding affinity of the EJC. Involved in translational enhancement of spliced mRNAs after formation of the 80S ribosome complex. Binds spliced mRNA in sequence-independent manner, 20-24 nucleotides upstream of mRNA exon-exon junctions. Shows higher affinity for single-stranded RNA in an ATP-bound core EJC complex than after the ATP is hydrolyzed.
- Tissue specificityUbiquitously expressed.
- Sequence similaritiesBelongs to the DEAD box helicase family. eIF4A subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
- Cellular localizationNucleus. Nucleus speckle. Cytoplasm. Nucleocytoplasmic shuttling protein. Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA. Detected in dendritic layer as well as the nuclear and cytoplasmic (somatic) compartments of neurons. Colocalizes with STAU1 and FMR1 in dendrites.
- Entrez Gene: 416704 Chicken
- Entrez Gene: 515145 Cow
- Entrez Gene: 9775 Human
- Entrez Gene: 192170 Mouse
- Entrez Gene: 688288 Rat
- Entrez Gene: 399362 Xenopus laevis
- Entrez Gene: 394053 Zebrafish
- Omim: 608546 Human
- SwissProt: Q5ZM36 Chicken
- SwissProt: Q2NL22 Cow
- SwissProt: P38919 Human
- SwissProt: Q91VC3 Mouse
- SwissProt: Q3B8Q2 Rat
- SwissProt: O42226 Xenopus laevis
- SwissProt: Q5U526 Xenopus laevis
- SwissProt: Q7ZVA6 Zebrafish
- Unigene: 389649 Human
- Unigene: 197555 Mouse
- Unigene: 422975 Mouse
- Unigene: 202617 Rat
- Unigene: 78203 Zebrafish
- ATP-dependent RNA helicase DDX48 antibodyATP-dependent RNA helicase eIF4A-3 antibodyDDX48 antibody
- DEAD box protein 48 antibodyeIF-4A-III antibodyeIF4A-III antibodyEIF4A3 antibodyeIF4AIII antibodyEukaryotic initiation factor 4A-III antibodyEukaryotic initiation factor 4A-like NUK-34 antibodyEUKARYOTIC TRANSLATION INITIATION FACTOR 4A ISOFORM 3 antibodyhNMP 265 antibodyIF4A3_HUMAN antibodyNMP 265 antibodyNMP265 antibodyNUCLEAR MATRIX PROTEIN 265 antibodyNUK34 antibody
Anti-eIF4A3 antibody images
eIF4A3 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to eIF4A3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32485.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 47kDa; eIF4A3
Anti-eIF4A3 antibody (ab32485) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg
IR Dye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size : 47 kDa
Observed band size : 47 kDa
All lanes : Anti-eIF4A3 antibody (ab32485) at 1 µg/ml
Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 47 kDa
Observed band size : 46 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab32485 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32485, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
IHC image of eIF4A3 staining in human skin FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32485, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
References for Anti-eIF4A3 antibody (ab32485)
This product has been referenced in:
- Chuang TW et al. The RNA-binding protein Y14 inhibits mRNA decapping and modulates processing body formation. Mol Biol Cell 24:1-13 (2013). WB, ICC/IF ; Human . Read more (PubMed: 23115303) »
- Feng J et al. A proteomic comparison of immature and mature mouse gonadotrophs reveals novel differentially expressed nuclear proteins that regulate gonadotropin gene transcription and RNA splicing. Biol Reprod 79:546-61 (2008). WB ; Mouse . Read more (PubMed: 18480465) »