Anti-eIF4A3 antibody (ab32485)
Key features and details
- Rabbit polyclonal to eIF4A3
- Suitable for: IHC-P, IP, ICC/IF, WB
- Reacts with: Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-eIF4A3 antibody
See all eIF4A3 primary antibodies -
Description
Rabbit polyclonal to eIF4A3 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Chicken, Cow, Xenopus laevis, Zebrafish, Cynomolgus monkey -
Immunogen
Synthetic peptide corresponding to Human eIF4A3 aa 1-100.
(Peptide available asab32484) -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP | (1) |
Use a concentration of 5 µg/ml.
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ICC/IF | (2) |
Use a concentration of 5 µg/ml.
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WB | (6) |
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
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Notes |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use a concentration of 5 µg/ml. |
ICC/IF
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa). |
Target
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Function
ATP-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 specificity
Ubiquitously expressed. -
Sequence similarities
Belongs to the DEAD box helicase family. eIF4A subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain. -
Cellular localization
Nucleus. 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. - Information by UniProt
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Database links
- Entrez Gene: 416704 Chicken
- Entrez Gene: 515145 Cow
- Entrez Gene: 101867456 Cynomolgus monkey
- Entrez Gene: 9775 Human
- Entrez Gene: 688288 Rat
- Entrez Gene: 399362 Xenopus laevis
- Entrez Gene: 394053 Zebrafish
- Omim: 608546 Human
see all -
Alternative names
- ATP-dependent RNA helicase DDX48 antibody
- ATP-dependent RNA helicase eIF4A-3 antibody
- DDX48 antibody
see all
Images
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Anti-eIF4A3 antibody (ab32485) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg
Secondary
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 -
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
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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.
Secondary
All lanes : 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).
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (17)
ab32485 has been referenced in 17 publications.
- Wang B et al. Eukaryotic initiation factor 4A3 inhibits Wnt/β-catenin signaling and regulates axis formation in zebrafish embryos. Development 148:N/A (2021). PubMed: 33914867
- Feng ZH et al. EIF4A3-induced circular RNA PRKAR1B promotes osteosarcoma progression by miR-361-3p-mediated induction of FZD4 expression. Cell Death Dis 12:1025 (2021). PubMed: 34716310
- Chen D et al. The circRAB3IP Mediated by eIF4A3 and LEF1 Contributes to Enzalutamide Resistance in Prostate Cancer by Targeting miR-133a-3p/miR-133b/SGK1 Pathway. Front Oncol 11:752573 (2021). PubMed: 34868959
- Herdlevaer I et al. Localization of CDR2L and CDR2 in paraneoplastic cerebellar degeneration. Ann Clin Transl Neurol 7:2231-2242 (2020). PubMed: 33009713
- Wang L et al. Circular RNA circSEMA5A promotes bladder cancer progression by upregulating ENO1 and SEMA5A expression. Aging (Albany NY) 12:21674-21686 (2020). PubMed: 33176280