Validated using a knockout cell line
Recombinant
RabMAb

Anti-eIF2A antibody [EPR11042] (ab169528)

Overview

  • Product name
    Anti-eIF2A antibody [EPR11042]
    See all eIF2A primary antibodies
  • Description
    Rabbit monoclonal [EPR11042] to eIF2A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human eIF2A aa 450-550.

  • Positive control
    • Molt4, Ramos, and BxPC-3 cell lysates; HeLa cells, PC-12 whole cell lysates, NIH/3T3 whole cell lysates; Human pancreas, Human prostatic hyperplasia, Human lung cancer, Mouse cerebrum, and Rat cerebrum tissues.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab169528 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 1/1000 - 1/5000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/50 - 1/100.

See IHC antigen retrieval protocols.

ICC/IF 1/250.

For unpurified use at 1/50 - 1/100.

  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function
      Functions in the early steps of protein synthesis of a small number of specific mRNAs. Acts by directing the binding of methionyl-tRNAi to 40S ribosomal subunits. In contrast to the eIF-2 complex, it binds methionyl-tRNAi to 40 S subunits in a codon-dependent manner, whereas the eIF-2 complex binds methionyl-tRNAi to 40 S subunits in a GTP-dependent manner. May act by impiging the expression of specific proteins.
    • Tissue specificity
      Widely expressed. Expressed at higher level in pancreas, heart, brain and placenta.
    • Sequence similarities
      Belongs to the WD repeat EIF2A family.
      Contains 3 WD repeats.
    • Information by UniProt
    • Database links
    • Alternative names
      • 65 kDa eukaryotic translation initiation factor 2A antibody
      • CDA 02 antibody
      • CDA02 antibody
      • EIF 2 antibody
      • EIF 2A antibody
      • eIF-2A antibody
      • EIF2 antibody
      • eif2a antibody
      • EIF2A_HUMAN antibody
      • Eukaryotic translation initiation factor 2A 65kDa antibody
      • Eukaryotic translation initiation factor 2A antibody
      • MST089 antibody
      • MSTP004 antibody
      • MSTP089 antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: eIF2A knockout HAP1 cell lysate (20 µg)
      Lane 3: Ramos cell lysate (20 µg)
      Lane 4: MOLT4 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab169528 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab169528 was shown to specifically react with eIF2A when eIF2A knockout samples were used. Wild-type and eIF2A knockout samples were subjected to SDS-PAGE. ab169528 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling eIF2A with purified ab169528 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    • All lanes : Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution (Unpurified)

      Lane 1 : Molt4 cell lysate
      Lane 2 : Ramos cell lysate
      Lane 3 : BxPC-3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 65 kDa
      Observed band size: 65 kDa

    • Immunohistochemical analysis of paraffin embedded Human pancreas tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.

    • Immunohistochemical analysis of paraffin embedded Human prostatic hyperplasia tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.

    References

    This product has been referenced in:
    • Meng C  et al. PERK Pathway Activation Promotes Intracerebral Hemorrhage Induced Secondary Brain Injury by Inducing Neuronal Apoptosis Both in Vivo and in Vitro. Front Neurosci 12:111 (2018). Read more (PubMed: 29541018) »

    See 1 Publication for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MDA231 cells)
    Gel Running Conditions
    Reduced Denaturing (10% Tris-Glycine Gel)
    Loading amount
    50000 cells
    Specification
    MDA231 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Aug 16 2016

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

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