Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-eEF1B2 antibody (ab37725)

Key features and details

  • Rabbit polyclonal to eEF1B2
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-eEF1B2 antibody
    See all eEF1B2 primary antibodies

  • Description

    Rabbit polyclonal to eEF1B2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human eEF1B2.

    Read Abcam's proprietary immunogen policy (Peptide available as ab37955.)

  • Positive control

    • HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate, Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate, A431 (Human epithelial carcinoma cell line) Whole Cell Lysate, HEK 293 (Human embryonic kidney cell line) Whole Cell Lysate, HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate, MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate and U2OS (Human osteosarcoma cell line) Whole Cell Lysate.

Properties

  • Form

    Liquid

  • Storage instructions

    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.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab37725 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
All applications
Mouse
Rabbit
Cow
Pig
Application Abreviews Notes
WB
1/250. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).
IHC-P
Use a concentration of 1 µg/ml.
ICC/IF
Use a concentration of 5 µg/ml.
Notes
WB
1/250. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).
IHC-P
Use a concentration of 1 µg/ml.
ICC/IF
Use a concentration of 5 µg/ml.

Target

  • Function

    EF-1-beta and EF-1-delta stimulate the exchange of GDP bound to EF-1-alpha to GTP.

  • Sequence similarities

    Belongs to the EF-1-beta/EF-1-delta family.
    Contains 1 GST C-terminal domain.

  • Post-translational
    modifications

    Phosphorylation affects the GDP/GTP exchange rate.
  • Information by UniProt

  • Database links

    • Alternative names

      • EEF1B antibody
      • EEF1B1 antibody
      • EEF1B2 antibody
      • EF-1-beta antibody
      • EF1B antibody
      • EF1B_HUMAN antibody
      • Elongation factor 1, beta-2-A antibody
      • Elongation factor 1-beta antibody
      • eukaryotic translation elongation factor 1 beta 1 antibody
      • eukaryotic translation elongation factor 1 beta 2 antibody
      see all

    Images

    • Western blot - Anti-eEF1B2 antibody (ab37725)
      Western blot - Anti-eEF1B2 antibody (ab37725)
      All lanes : Anti-eEF1B2 antibody (ab37725) at 1/500 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : EEF1B2 knockout HEK293T cell lysate
      Lane 3 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 25 kDa
      Observed band size: 30 kDa
      why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab37725 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.

       ab37725 Anti-eEF1B2 antibody was shown to specifically react with eEF1B2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266769 (knockout cell lysate ab257934) was used. Wild-type and eEF1B2 knockout samples were subjected to SDS-PAGE. ab37725 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-eEF1B2 antibody (ab37725)
      Immunocytochemistry/ Immunofluorescence - Anti-eEF1B2 antibody (ab37725)
      ICC/IF image of ab37725 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37725, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and Hek293 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eEF1B2 antibody (ab37725)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eEF1B2 antibody (ab37725)
      IHC image of ab37725 staining eEF1B2 in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37725, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    • Western blot - Anti-eEF1B2 antibody (ab37725)
      Western blot - Anti-eEF1B2 antibody (ab37725)
      All lanes : Anti-eEF1B2 antibody (ab37725) at 1/250 dilution

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat whole cell lysate (ab7899)
      Lane 3 : A-431 whole cell lysate (ab7909)
      Lane 4 : HEK-293 whole cell lysate (ab7902)
      Lane 5 : Hep G2 whole cell lysate (ab7900)
      Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
      Lane 7 : U2OS (Human osteosarcoma 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 1/15000 dilution

      Performed under reducing conditions.

      Predicted band size: 25 kDa
      Observed band size: 30 kDa why is the actual band size different from the predicted?

    References (3)

    Publishing research using ab37725? Please let us know so that we can cite the reference in this datasheet.

    ab37725 has been referenced in 3 publications.

    • Jia L  et al. Translation elongation factor eEF1Ba is identified as a novel prognostic marker of gastric cancer. Int J Biol Macromol 126:345-351 (2018). PubMed: 30572058
    • Sivan G  et al. Mitotic modulation of translation elongation factor 1 leads to hindered tRNA delivery to ribosomes. J Biol Chem 286:27927-35 (2011). WB ; Human . PubMed: 21665947
    • Byun HO  et al. Cathepsin D and eukaryotic translation elongation factor 1 as promising markers of cellular senescence. Cancer Res 69:4638-47 (2009). WB ; Human . PubMed: 19487283

    Images

    • Western blot - Anti-eEF1B2 antibody (ab37725)
      Western blot - Anti-eEF1B2 antibody (ab37725)
      All lanes : Anti-eEF1B2 antibody (ab37725) at 1/500 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : EEF1B2 knockout HEK293T cell lysate
      Lane 3 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 25 kDa
      Observed band size: 30 kDa
      why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab37725 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.

       ab37725 Anti-eEF1B2 antibody was shown to specifically react with eEF1B2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266769 (knockout cell lysate ab257934) was used. Wild-type and eEF1B2 knockout samples were subjected to SDS-PAGE. ab37725 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-eEF1B2 antibody (ab37725)
      Immunocytochemistry/ Immunofluorescence - Anti-eEF1B2 antibody (ab37725)
      ICC/IF image of ab37725 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37725, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and Hek293 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eEF1B2 antibody (ab37725)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eEF1B2 antibody (ab37725)
      IHC image of ab37725 staining eEF1B2 in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37725, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    • Western blot - Anti-eEF1B2 antibody (ab37725)
      Western blot - Anti-eEF1B2 antibody (ab37725)
      All lanes : Anti-eEF1B2 antibody (ab37725) at 1/250 dilution

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat whole cell lysate (ab7899)
      Lane 3 : A-431 whole cell lysate (ab7909)
      Lane 4 : HEK-293 whole cell lysate (ab7902)
      Lane 5 : Hep G2 whole cell lysate (ab7900)
      Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
      Lane 7 : U2OS (Human osteosarcoma 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 1/15000 dilution

      Performed under reducing conditions.

      Predicted band size: 25 kDa
      Observed band size: 30 kDa why is the actual band size different from the predicted?

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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