RabMAb

Anti-EEF2抗体[EP880Y] (ab75748)

概述

  • 产品名称Anti-EEF2抗体[EP880Y]
    参阅全部 EEF2 一抗
  • 描述
    兔单克隆抗体[EP880Y] to EEF2
  • 经测试应用适用于: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues on the C terminal of human EEF2.

  • 阳性对照
    • A431, NIH3T3, HeLa, C6, PC-12 and NIH/3T3 cell lysates. Kidney carcinoma tissue, rat pancreas, mouse pancreas, human glioma.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    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.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.1% BSA
  • Concentration information loading...
  • 纯度Protein A purified
  • 克隆单克隆
  • 克隆编号EP880Y
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab75748 in the following tested applications.

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

应用 Ab评论 说明
WB 1/10000 - 1/50000. Detects a band of approximately 95 kDa (predicted molecular weight: 95 kDa).

 

IP 1/20.
Flow Cyt 1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.

靶标

  • 功能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.
  • 序列相似性Belongs to the GTP-binding elongation factor family. EF-G/EF-2 subfamily.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-EEF2 antibody [EP880Y] 图像

  • All lanes : Anti-EEF2 antibody [EP880Y] (ab75748) at 1/50000 dilution

    Lane 1 : C6 (rat glioma) whole cell lysates
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 3 : NIH/3T3 (mouse embryo) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Rabbit monoclonal [EP880Y] to EEF2 (ab75748) at 1/100000 dilution

    Predicted band size : 95 kDa
  • ab75748 staining EEF2 in rat pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • ab75748 staining EEF1 in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab75748 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

  • ab75748 staining EEF2 in the human cell line HEK293 (human embryonic kidney) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/70. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • ab75748 immunoprecipitating EEF2. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
    Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab75748 in HeLa (human cervix adenocarcinoma) whole cell lysate

  • All lanes : Anti-EEF2 antibody [EP880Y] (ab75748) at 1/50000 dilution

    Lane 1 : HEK293 (human embryonic kidney) whole cell lysates
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysates
    Lane 3 : A431 (human epidermoid carcinoma) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 95 kDa
    Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.
  • ab75748 staining EEF2 in mouse pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

    Negative control 1: PBS in place of primary antibody.

  • Overlay histogram showing HeLa cells stained with ab75748 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75748, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • EEF2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to EEF2 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 ab75748.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 95kDa; EEF2
  • ab75748 staining EEF2 in human glioma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

    Negative control 1: PBS in place of primary antibody.

Anti-EEF2 antibody [EP880Y] (ab75748)参考文献

ab75748 has not yet been referenced specifically in any publications.

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