Overview

  • Product name

    Anti-EEF2/Elongation factor 2 antibody
    See all EEF2/Elongation factor 2 primary antibodies
  • Description

    Rabbit polyclonal to EEF2/Elongation factor 2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chicken, Hamster, Cow, Xenopus laevis
  • Immunogen

    Synthetic peptide corresponding to Human EEF2/Elongation factor 2 aa 400-500 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab33522)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; HEK 293; HepG2; MCF-7; SHSY-5Y; U2OS; NIH 3T3; MEF1; PC12. This antibody gave a positive signal in the following tissue lysates: Mouse Pancreas; Mouse Ovary; Rat Liver. IHC-P: human lung FFPE tissue sections
  • General notes

     This product was previously labelled as EEF2

     

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

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab33523 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
IP Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Detects a band of approximately 95 kDa (predicted molecular weight: 95 kDa).Can be blocked with Human EEF2/Elongation factor 2 peptide (ab33522).

Target

  • Function

    Catalyzes the GTP-dependent ribosomal translocation step during translation elongation. During this step, the ribosome changes from the pre-translocational (PRE) to the post-translocational (POST) state as the newly formed A-site-bound peptidyl-tRNA and P-site-bound deacylated tRNA move to the P and E sites, respectively. Catalyzes the coordinated movement of the two tRNA molecules, the mRNA and conformational changes in the ribosome.
  • Sequence similarities

    Belongs to the GTP-binding elongation factor family. EF-G/EF-2 subfamily.
  • Post-translational
    modifications

    Phosphorylation by EF-2 kinase completely inactivates EF-2.
    Diphthamide is 2-[3-carboxyamido-3-(trimethyl-ammonio)propyl]histidine. Diphthamide can be ADP-ribosylated by diphtheria toxin and by Pseudomonas exotoxin A, thus arresting protein synthesis.
    ISGylated.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • EEF 2 antibody
    • Eef2 antibody
    • EF-2 antibody
    • EF2 antibody
    • EF2_HUMAN antibody
    • Elongation factor 2 antibody
    • Eukaryotic translation elongation factor 2 antibody
    • Polypeptidyl tRNA translocase antibody
    • SCA26 antibody
    see all

Images

  • All lanes : Anti-EEF2/Elongation factor 2 antibody (ab33523) 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)
    Lane 4 : HEK293 whole cell lysate (ab7902)
    Lane 5 : HepG2 whole cell lysate (ab7900)
    Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human Neuroblastoma) Whole Cell Lysate
    Lane 8 : 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: 95 kDa
    Observed band size: 95 kDa

  • All lanes : Anti-EEF2/Elongation factor 2 antibody (ab33523) 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 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 4 : Ovary (Mouse) Tissue Lysate - normal tissue
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 6 : Liver (Rat) Tissue Lysate - normal tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size: 95 kDa
    Observed band size: 95 kDa

  • ICC/IF image of ab33523 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33523, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

  • All lanes : Anti-EEF2/Elongation factor 2 antibody (ab33523) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate (20ug)
    Lane 3 : Zebrafish heart homogenate (20ug)
    Lane 4 : Zebrafish liver homogenate (20ug)
    Lane 5 : Zebrafish skeletal muscle homogenate (20ug)
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (20ug)

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 95 kDa
    Observed band size: 95 kDa


    Exposure time: 5 minutes
  • IHC image of EEF2/Elongation factor 2 staining in human lung 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 ab33523, 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.

  • EEF2/Elongation factor 2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to EEF2/Elongation factor 2 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 ab33523.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 95kDa; EEF2/Elongation factor 2, non-specific band: 135kDa,due to background seen in No ab control lane (2). 

References

This product has been referenced in:

  • Strigli A  et al. Doxorubicin induces caspase-mediated proteolysis of KV7.1. Commun Biol 1:155 (2018). Read more (PubMed: 30302399) »
  • Kissing S  et al. Disruption of the vacuolar-type H+-ATPase complex in liver causes MTORC1-independent accumulation of autophagic vacuoles and lysosomes. Autophagy 13:670-685 (2017). Read more (PubMed: 28129027) »
See all 7 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3 cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
3T3 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 12 2007

Application
Western blot
Sample
Human Cell lysate - whole cell (293T, U2OS and MCF7 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
293T, U2OS and MCF7 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 12 2007

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