Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-eIF2A antibody [EPR11042] (ab169528)

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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

    • WB: Molt4, Ramos, HAP1, PC-12, and NIH/3T3 lysates. IHC-P: Human pancreas, Human prostatic hyperplasia, Human lung cancer, Mouse cerebrum, and Rat cerebrum tissues. ICC/IF: HeLa cells, MCF7 cells.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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.

Perform heat mediated antigen retrieval.

 
ICC/IF 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

      see all

    • 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

    • Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
      Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

      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 - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling eIF2A with Purified ab169528 at 1:100 dilution (4.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
      Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
      Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution (Purified) + NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

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

      Lane 1 : Ramos (Human Burkitt's lymphoma Blymphocyte) whole cell lysates
      Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

    • Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)

      ab169528 staining eIF2A in wild-type HAP1 cells (top panel) and EIF2A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab169528 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

      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) »
    • Zhang Z  et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). WB ; Mouse . Read more (PubMed: 30297870) »
    See all 4 Publications for this product

    Publishing research using ab169528? Please let us know so that we can cite the reference in this datasheet.

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    Western blot abreview for Anti-eIF2A antibody [EPR11042]

     Excellent
    Abreviews
    abreview image
    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%
    Read More

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    Verified customer

    Submitted Aug 16 2016

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