Overview

  • Product name
    Anti-eIF4A1 antibody
    See all eIF4A1 primary antibodies
  • Description
    Rabbit polyclonal to eIF4A1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, African green monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human eIF4A1.

    Read Abcam's proprietary immunogen policy

    (Peptide available as ab31796.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) This antibody gave a positive signal in the following tissue lysates: Liver (Mouse) Pancreas (Mouse) Testis (Mouse)

Properties

Applications

Our Abpromise guarantee covers the use of ab31217 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the DEAD box helicase family. eIF4A subfamily.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • ATP-dependent RNA helicase eIF4A-1 antibody
    • DDX2 antibody
    • DDX2A antibody
    • eIF 4A I antibody
    • eIF-4A-I antibody
    • EIF4A antibody
    • eIF4A I antibody
    • eIF4A-I antibody
    • EIF4A1 antibody
    • Eukaryotic initiation factor 4A-I antibody
    • Eukaryotic initiation factor 4AI antibody
    • eukaryotic translation initiation factor 4A, isoform 1 antibody
    • Eukaryotic translation initiation factor 4A1 antibody
    • IF4A1_HUMAN antibody
    see all

Images

  • Western blot - Anti-eIF4A1 antibody (ab31217)
    Western blot - Anti-eIF4A1 antibody (ab31217)
    All lanes : Anti-eIF4A1 antibody (ab31217) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4A1 antibody (ab31217)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4A1 antibody (ab31217)
    ICC/IF image of ab31217 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab31217, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence.  5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4A1 antibody (ab31217)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4A1 antibody (ab31217)

    Image courtesy of Human Protein Atlas. ab31217 staining eIF4A1 in human stomach, showing strong cytoplasmic staining in the glandular cells. Paraffin embedded human stomach tissue was incubated with ab31217 (1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab31217 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

  • Western blot - Anti-eIF4A1 antibody (ab31217)
    Western blot - Anti-eIF4A1 antibody (ab31217)
    All lanes : Anti-eIF4A1 antibody (ab31217) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 4 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 5 : Testis (Mouse) Tissue Lysate - normal tissue
    Lane 6 : 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: 47 kDa

  • Immunoprecipitation - Anti-eIF4A1 antibody (ab31217)
    Immunoprecipitation - Anti-eIF4A1 antibody (ab31217)
    eIF4A1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to eIF4A1 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31217.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 47kDa: eIF4A1

References

This product has been referenced in:
  • Amorim IS  et al. Loss of eIF4E Phosphorylation Engenders Depression-like Behaviors via Selective mRNA Translation. J Neurosci 38:2118-2133 (2018). Read more (PubMed: 29367404) »
  • Passacantilli I  et al. Alternative polyadenylation of ZEB1 promotes its translation during genotoxic stress in pancreatic cancer cells. Cell Death Dis 8:e3168 (2017). WB . Read more (PubMed: 29120411) »

See all 24 Publications for this product

Publishing research using ab31217? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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7 reviews or Q&A

10
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Western blot abreview for Anti-eIF4A1 antibody

 Excellent
Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells (J1))
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
mouse embryonic stem cells (J1)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Feb 01 2018

Western blot abreview for Anti-eIF4A1 antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse neuroblastoma N2A cells)
Loading amount
30 µg
Specification
Mouse neuroblastoma N2A cells
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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Submitted Jul 03 2012

Western blot abreview for Anti-eIF4A1 antibody

 Excellent
Abreviews
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
20 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
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Submitted Jun 02 2012

Immunoprecipitation abreview for Anti-eIF4A1 antibody

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - other (SW480)
Total protein in input
30 µg
Specification
SW480
Immuno-precipitation step
Protein G
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Submitted May 24 2012

Western blot abreview for Anti-eIF4A1 antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Vero Cells)
Loading amount
10 µg
Specification
Vero Cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Oct 24 2007

Immunocytochemistry/ Immunofluorescence abreview for Anti-eIF4A1 antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% NP40
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
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Submitted Oct 23 2007

I would like to have informations about your eIF4A1 antibody (ab 31217). I use the sea urchin embryos for my analysis. I would like to know the sequence similarity and identity between the sea urchin eIF4A and the peptide that you used to produce thi...

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Thank you for your enquiry. Unfortunately it appears there is no significant homology between the immunogen and the seuence you provided. It is unlikely this antibody will cross-react with sea urchin. I hope this information helps, please do not...

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