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Epigenetics and Nuclear Signaling DNA / RNA Translation Regulation

Anti-eIF4A3抗体(ab32485)

  • Datasheet
  • SDS
Reviews (9)Q&A (1)References (21)

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Western blot - Anti-eIF4A3 antibody (ab32485)
  • Immunoprecipitation - Anti-eIF4A3 antibody (ab32485)
  • Western blot - Anti-eIF4A3 antibody (ab32485)
  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4A3 antibody (ab32485)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

选择批间可重复性更高的重组抗体

Product image
Anti-eIF4A3 antibody [EPR14301(B)] (ab180573)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-eIF4A3抗体
    参阅全部 eIF4A3 一抗
  • 描述

    兔多克隆抗体to eIF4A3
  • 宿主

    Rabbit
  • 经测试应用

    适用于: IHC-P, IP, ICC/IF, WBmore details
  • 种属反应性

    与反应: Rat, Human
    预测可用于: Chicken, Cow, Xenopus laevis, Zebrafish, Cynomolgus monkey
  • 免疫原

    Synthetic peptide corresponding to Human eIF4A3 aa 1-100.
    (Peptide available as ab32484)

  • 常规说明

    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

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 98.98% PBS, 1% BSA

    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...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Regulation

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Human eIF4A3 protein (ab123151)
  • Recombinant Protein

    • Recombinant Human eIF4A3 protein (ab123151)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab32485于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

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.
IP (1)
Use a concentration of 5 µg/ml.
ICC/IF (2)
Use a concentration of 5 µg/ml.
WB (6)
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
说明
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).

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.
  • Target information above from: UniProt accession P38919 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • 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
    • SwissProt: Q5ZM36 Chicken
    • SwissProt: Q2NL22 Cow
    • SwissProt: Q4R3Q1 Cynomolgus monkey
    • SwissProt: P38919 Human
    • SwissProt: Q3B8Q2 Rat
    • SwissProt: O42226 Xenopus laevis
    • SwissProt: Q7ZVA6 Zebrafish
    • Unigene: 389649 Human
    • Unigene: 202617 Rat
    • Unigene: 78203 Zebrafish
    see all
  • 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

图片

  • Western blot - Anti-eIF4A3 antibody (ab32485)
    Western blot - Anti-eIF4A3 antibody (ab32485)
    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

  • Immunoprecipitation - Anti-eIF4A3 antibody (ab32485)
    Immunoprecipitation - Anti-eIF4A3 antibody (ab32485)

    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

  • Western blot - Anti-eIF4A3 antibody (ab32485)
    Western blot - Anti-eIF4A3 antibody (ab32485)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4A3 antibody (ab32485)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4A3 antibody (ab32485)
    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4A3 antibody (ab32485)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4A3 antibody (ab32485)
    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.

实验方案

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (21)

发表研究结果有使用 ab32485?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab32485 被引用在 21 文献中.

  • Hu G  et al. Exosomal circCOL1A1 promotes angiogenesis via recruiting EIF4A3 protein and activating Smad2/3 pathway in colorectal cancer. Mol Med 29:155 (2023). PubMed: 37940881
  • Lehtiniemi T  et al. SMG6 localizes to the chromatoid body and shapes the male germ cell transcriptome to drive spermatogenesis. Nucleic Acids Res 50:11470-11491 (2022). PubMed: 36259644
  • Ma J  et al. Circ 003390/Eukaryotic translation initiation factor 4A3 promoted cell migration and proliferation in endometrial cancer via vascular endothelial growth factor signaling by miR-195-5p. Bioengineered 13:11958-11972 (2022). PubMed: 35546509
  • 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
View all Publications for this product

客户评价及客户问答

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1-10 of 10 Abreviews or Q&A

Western blot abreview for Anti-eIF4A3 antibody

Good
Abreviews
Abreviews
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 10 2017

Immunoprecipitation abreview for Anti-eIF4A3 antibody

Below Average
Abreviews
Abreviews
abreview image
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)
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Jianlong Wang

Verified customer

提交于 Jan 31 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-eIF4A3 antibody

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jan 30 2017

Western blot abreview for Anti-eIF4A3 antibody

Poor
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jan 27 2017

Western blot

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 07 2013

Immunocytochemistry/ Immunofluorescence

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Loredana Bury

Verified customer

提交于 May 31 2013

Western blot abreview for Anti-eIF4A3 antibody

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jul 29 2011

Western blot abreview for Anti-eIF4A3 antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jul 07 2011

Western blot abreview for Anti-eIF4A3 antibody

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 01 2008

Question

Very weak signal and high background in WB

Read More

Abcam community

Verified customer

Asked on Oct 16 2012

Answer

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 protocol tips that we discussed do not prove to be successful, then please let me know and I would be happy to either send a replacement antibody or to process a refund.

If there is anything else I can help you with, please let me know.

Read More

Abcam Scientific Support

回复于 Oct 16 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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