概述

  • 产品名称
  • 描述
    兔多克隆抗体to eIF4A3
  • 经测试应用
    适用于: IHC-P, IHC-Fr, IP, ICC/IF, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Chicken, Cow, Xenopus laevis, Zebrafish, Cynomolgus monkey
  • 免疫原

    Synthetic peptide derived from within residues 1 - 100 of Human eIF4A3.

    (Peptide available as ab32484.)

  • 阳性对照
    • This 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

性能

  • 形式
    Liquid
  • 存放说明
    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.
  • 存储溶液
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS. pH 7.4
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

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.

应用 Ab评论 说明
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration. PubMed: 22982623
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).

靶标

  • 功能
    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.
  • 组织特异性
    Ubiquitously expressed.
  • 序列相似性
    Belongs to the DEAD box helicase family. eIF4A subfamily.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
  • 细胞定位
    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
  • 数据库链接
  • 别名
    • ATP-dependent RNA helicase DDX48 antibody
    • ATP-dependent RNA helicase eIF4A-3 antibody
    • DDX48 antibody
    • DEAD box protein 48 antibody
    • eIF-4A-III antibody
    • eIF4A-III antibody
    • EIF4A3 antibody
    • eIF4AIII antibody
    • Eukaryotic initiation factor 4A-III antibody
    • Eukaryotic initiation factor 4A-like NUK-34 antibody
    • Eukaryotic translation initiation factor 4A isoform 3 antibody
    • hNMP 265 antibody
    • IF4A3_HUMAN antibody
    • NMP 265 antibody
    • NMP265 antibody
    • Nuclear matrix protein 265 antibody
    • NUK34 antibody
    see all

Anti-eIF4A3 antibody 图像

  • 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

    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
  • 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
    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.

Anti-eIF4A3 antibody (ab32485)参考文献

This product has been referenced in:
  • Van Rechem C  et al. Lysine demethylase KDM4A associates with translation machinery and regulates protein synthesis. Cancer Discov 5:255-63 (2015). WB ; Human . Read more (PubMed: 25564516) »
  • 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) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Bis-Tris)
Loading amount
15 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Apr 10 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Mouse embryonic stem cell line (CCE))
Total protein in input
6e+006 cells
Immuno-precipitation step
Protein G
Specification
Mouse embryonic stem cell line (CCE)
Username

Jianlong Wang

Verified customer

提交于 Jan 31 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse ESC (CCE))
Permeabilization
Yes - 0.25% TritonX-100 solution (dilute in PBS)
Specification
mouse ESC (CCE)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jan 30 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse embryonic stem cell line (CCE))
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Mouse embryonic stem cell line (CCE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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提交于 Jan 27 2017

Western blot

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Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (Megakaryocytes)
Specification
Megakaryocytes
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

提交于 Jun 07 2013

Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Chinese Hamster Cell (CHO cell line)
Specification
CHO cell line
Permeabilization
Yes - Triton-X 0.1%
Fixative
Paraformaldehyde
Username

Dr. Loredana Bury

Verified customer

提交于 May 31 2013

Thank you for contacting Abcam.

I am sorry about the issues you have been having with ab32485 in western blot. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples . If the prot...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse embryonic Fibroblast)
Loading amount
25 µg
Specification
Mouse embryonic Fibroblast
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jul 29 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
15 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jul 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 (20 ug) and HeLa (40 ug))
Loading amount
40 µg
Specification
HEK293 (20 ug) and HeLa (40 ug)
Gel Running Conditions
Reduced Denaturing (10 %)
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 May 01 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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