製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to eIF4A2
  • アプリケーション
    適用あり: IHC-P, IP, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human eIF4A2.

    (Peptide available as ab32473.)

  • ポジティブ・コントロール
    • Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Brain (Mouse) Tissue Lysate Testis (Mouse) Tissue Lysate Skeletal Muscle (Mouse) Tissue Lysate (ab29711) Spinal Cord (Mouse) Tissue Lysate Ovary (Mouse) Tissue Lysate PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Brain (Rat) Tissue Lysate Heart (Rat) Tissue Lysate

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    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.
  • バッファー
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab31218 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
ICC/IF 1/200.

ターゲット情報

  • 機能
    ATP-dependent RNA helicase which is a subunit of the eIF4F complex involved in cap recognition and is required for mRNA binding to ribosome. In the current model of translation initiation, eIF4A unwinds RNA secondary structures in the 5'-UTR of mRNAs which is necessary to allow efficient binding of the small ribosomal subunit, and subsequent scanning for the initiator codon.
  • 配列類似性
    Belongs to the DEAD box helicase family. eIF4A subfamily.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
  • Information by UniProt
  • 参照データベース
  • 別名
    • ATP dependent RNA helicase eIF4A 2 antibody
    • ATP-dependent RNA helicase eIF4A-2 antibody
    • BM-010 antibody
    • DDX2B antibody
    • eIF 4A II antibody
    • eIF-4A-II antibody
    • EIF4A antibody
    • eIF4A II antibody
    • eIF4A-II antibody
    • EIF4A2 antibody
    • EIF4F antibody
    • Eukaryotic initiation factor 4A II antibody
    • Eukaryotic initiation factor 4A-II antibody
    • eukaryotic translation initiation factor 4A isoform 2 antibody
    • Eukaryotic translation initiation factor 4A2 antibody
    • IF4A2_HUMAN antibody
    • N-terminally processed antibody
    see all

画像

  • Anti-eIF4A2 antibody (ab31218) at 1 µg/ml + Jurkat whole cell lysate (ab7899) 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
    Additional bands at : 52 kDa. We are unsure as to the identity of these extra bands.
  • Immunocytochemistry/ Immunofluorescence analysis of MDA-MB231 Cells labeling eIF4A2 with ab31218 at a dilution of 1/200. Cells were fixed with Paraformaldehyde and permeabilized with 0.2% Saponin. The blocking step was incubation with 5% BSA for 30 minutes at 22°C. An Alexa Flour® 488 conjugated donkey anti-rabbit was used as the secondary antibody.

    See Abreview

  • Image courtesy of Human Protein Atlas. ab31218 staining eIF4A2 in human duodenum, showing strong cytomplasmic staining in the villi and Brunner's gland cells. Paraffin embedded human duodenum tissue was incubated with ab31218 (1/400 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab31218 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues. Further results for this antibody can be found at www.proteinatlas.org

  • eIF4A2 was immunoprecipitated using 0.5mg Mouse skeletal muscle whole tissue extract, 5µg of Rabbit polyclonal to eIF4A2 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, Mouse skeletal muscle whole tissue 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31218.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 47kDa: eIF4A2 .

  • All lanes : Anti-eIF4A2 antibody (ab31218) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Testis (Mouse) Tissue Lysate
    Lane 3 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 4 : Spinal Cord (Mouse) Tissue Lysate
    Lane 5 : Ovary (Mouse) Tissue Lysate
    Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 7 : Brain (Rat) Tissue Lysate
    Lane 8 : Heart (Rat) Tissue 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 : 47 kDa
    Additional bands at : 51 kDa. We are unsure as to the identity of these extra bands.

参考文献

This product has been referenced in:
  • Seo J  et al. Oxidative Stress Triggers Body-Wide Skipping of Multiple Exons of the Spinal Muscular Atrophy Gene. PLoS One 11:e0154390 (2016). WB . Read more (PubMed: 27111068) »
  • Xi S  et al. Downregulation of ring-finger protein 43 in glioma associates with poor prognosis. Int J Clin Exp Pathol 8:490-6 (2015). Read more (PubMed: 25755738) »

See all 13 Publications for this product

レビューと Q&A

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 cells (CCE))
Total protein in input
600000 cells
Immuno-precipitation step
Protein G
Specification
mouse embryonic stem cells (CCE)
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投稿 Jan 30 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse ES Cells (CCE))
Permeabilization
Yes - 0.25% TritonX-100 solution (dilute in PBS)
Specification
Mouse ES Cells (CCE)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
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投稿 Jan 27 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 cells (CCE))
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
shRNA knockdown
Specification
mouse embryonic stem cells (CCE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Jan 26 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (hek293, hela)
Loading amount
50 µg
Specification
hek293, 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
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投稿 Feb 25 2011

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 fibroblasts)
Loading amount
40 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (7.5)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Feb 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (MDA-MB231 and Hela Cell lines)
Loading amount
50000 cells
Specification
MDA-MB231 and Hela Cell lines
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Jul 05 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (MDA-MB231 Cells)
Specification
MDA-MB231 Cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.2% Saponin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Jul 04 2007

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

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