Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-EEF1A2 antibody [EPR22651-46] (ab227824)

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Overview

  • Product name

    Anti-EEF1A2 antibody [EPR22651-46]
    See all EEF1A2 primary antibodies

  • Description

    Rabbit monoclonal [EPR22651-46] to EEF1A2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human EEF1A2 aa 200-300. The exact sequence is proprietary.
    Database link: Q05639

  • Positive control

    • WB: Wild-type HAP1, HEK-293T, Neuro 2a whole cell lysates; Human brain tissue lysate; Mouse and rat brain and heart tissue lysate. IP: HEK-293T whole cell lysate.

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab227824 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. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

    • Target

    • Function

      This protein promotes the GTP-dependent binding of aminoacyl-tRNA to the A-site of ribosomes during protein biosynthesis.

    • Tissue specificity

      Brain, heart, and skeletal muscle.

    • Sequence similarities

      Belongs to the GTP-binding elongation factor family. EF-Tu/EF-1A subfamily.

    • Cellular localization

      Nucleus.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • eEF1A-2 antibody
      • Eef1a2 antibody
      • EEF1AL antibody
      • EF 1 alpha 2 antibody
      • EF-1-alpha-2 antibody
      • EF1A antibody
      • EF1A2_HUMAN antibody
      • Elongation factor 1 A 2 antibody
      • Elongation factor 1 alpha antibody
      • Elongation factor 1 alpha 2 antibody
      • Elongation factor 1-alpha 2 antibody
      • Eukaryotic elongation factor 1 A-2 antibody
      • Eukaryotic translation elongation factor 1 alpha 2 antibody
      • FLJ41696 antibody
      • HS1 antibody
      • OTTHUMP00000031776 antibody
      • Statin like antibody
      • Statin S1 antibody
      • Statin-S1 antibody
      • STN antibody
      • STNL antibody
      see all

    Images

    • Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      All lanes : Anti-EEF1A2 antibody [EPR22651-46] (ab227824) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : EEF1A2 knockout HAP1 whole cell lysate
      Lane 3 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
      Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 50 kDa
      Observed band size: 50 kDa



      ab227824 was shown to specifically react with EEF1A2 in wild-type HAP1 cells as signal was lost in EEF1A2 knockout cells. Wild-type and EEF1A2 knockout samples were subjected to SDS-PAGE. ab227824 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      This antibody also recognizes a weakly unidentifiable band at around 75kDa

      Blocking/diluting buffer and concentration: 5% NFDM/TBST

      Exposure time: 3.25 seconds

    • Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      Anti-EEF1A2 antibody [EPR22651-46] (ab227824) at 1/1000 dilution + Human brain tissue lysate at 20 µg

      Secondary
      VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 50 kDa
      Observed band size: 50 kDa



      Blocking/diluting buffer and concentration: 5% NFDM/TBST

      Exposure time: 7.75 seconds

    • Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      Western blot - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      All lanes : Anti-EEF1A2 antibody [EPR22651-46] (ab227824) at 1/1000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Rat brain tissue lysate
      Lane 4 : Rat heart tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 50 kDa
      Observed band size: 50 kDa



      Blocking/diluting buffer and concentration: 5% NFDM/TBST

      Exposure Time: 3.25 seconds

    • Immunoprecipitation - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)
      Immunoprecipitation - Anti-EEF1A2 antibody [EPR22651-46] (ab227824)

      EEF1A2 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate with ab227824 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Ab227824 1/1000 dilution (0.54 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

      Lane 1: HEK-293T (human embryonic kidney epithelial cell) whole cell lysate 10μg

      Lane 2: ab227824 IP in HEK-293T whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227824 in HEK-293T whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 15 seconds

    References

    ab227824 has not yet been referenced specifically in any publications.

    Publishing research using ab227824? Please let us know so that we can cite the reference in this datasheet.

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