Recombinant
RabMAb

Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

Overview

  • Product name
    Anti-ATF2 (phospho T71) antibody [E268]
    See all ATF2 primary antibodies
  • Description
    Rabbit monoclonal [E268] to ATF2 (phospho T71)
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
    Unsuitable for: Flow Cyt or IHC
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human ATF2 aa 50-150.
    Database link: P15336

  • Positive control
    • HeLa cells. NIH/3T3 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    SAPK and p38 MAPK activate, in response to cellular stress, ATF2 by phosphorylating the protein at Thr69 and Thr71. Mutations of these sites result in the loss of stress induced transcription by ATF2.

     

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

    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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • Purity
    Protein A purified
  • Primary antibody notes
    SAPK and p38 MAPK activate, in response to cellular stress, ATF2 by phosphorylating the protein at Thr69 and Thr71. Mutations of these sites result in the loss of stress induced transcription by ATF2.
  • Clonality
    Monoclonal
  • Clone number
    E268
  • Isotype
    IgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab32019 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 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 54 kDa).
IP 1/30.
ICC/IF 1/500.
  • Application notes
    Is unsuitable for Flow Cyt or IHC.
  • Target

    • Function
      Transcriptional activator, probably constitutive, which binds to the cAMP-responsive element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Interaction with JUN redirects JUN to bind to CRES preferentially over the 12-O-tetradecanoylphorbol-13-acetate response elements (TRES) as part of an ATF2/JUN complex.
    • Tissue specificity
      Abundant expression seen in the brain.
    • Sequence similarities
      Belongs to the bZIP family. ATF subfamily.
      Contains 1 bZIP domain.
      Contains 1 C2H2-type zinc finger.
    • Post-translational
      modifications
      Phosphorylation of Thr-69 and Thr-71 by MAPK14 causes increased transcriptional activity. Also phosphorylated and activated by JNK.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • Activating transcription factor 2 antibody
      • Activating transcription factor 2 splice variant ATF2 var2 antibody
      • ATF 2 antibody
      • Atf-2 antibody
      • Atf2 antibody
      • ATF2 protein antibody
      • ATF2_HUMAN antibody
      • cAMP Response Element Binding Protein 2 antibody
      • cAMP response element binding protein CRE BP1 antibody
      • cAMP response element-binding protein CRE-BP1 antibody
      • cAMP responsive element binding protein 2, formerly antibody
      • cAMP-dependent transcription factor ATF-2 antibody
      • cAMP-responsive element-binding protein 2 antibody
      • CRE BP1 antibody
      • CRE-BP antibody
      • CREB 2 antibody
      • CREB-2 antibody
      • CREB2 antibody
      • CREBP1 antibody
      • Cyclic AMP dependent transcription factor ATF 2 antibody
      • Cyclic AMP-dependent transcription factor ATF-2 antibody
      • Cyclic AMP-responsive element-binding protein 2 antibody
      • D130078H02Rik antibody
      • D18875 antibody
      • HB 16 antibody
      • HB16 antibody
      • Histone acetyltransferase ATF2 antibody
      • MGC105211 antibody
      • MGC105222 antibody
      • MGC111558 antibody
      • MGC142504 antibody
      • mXBP antibody
      • MXBP protein antibody
      • Tg(Gzma-Klra1)7Wum antibody
      • TREB 7 antibody
      • TREB7 antibody
      see all

    Anti-ATF2 (phospho T71) antibody [E268] images

    • Immunocytochemistry of HeLa (Human epithelial cell line from cervix adenocarcinoma), prepared in FBS free medium overnight labeling ATF2 at 0.9 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 1/500 . Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as the counter stain at 2.5 μg/ml. DAPI was used for nuclear counter stain. Confocal image showing the expression was increased on HeLa cells, prepared in FBS free medium overnight, then treated with 250ng/ml anisomycin for 30min.

       

       

    • All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.5 µg/ml

      Lane 1 : As above
      Lane 2 : As above

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

      Predicted band size : 54 kDa


      Exposure time : 3 minutes

      Blocking and diluting buffer used was 5% NFDM/TBST

    • All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.1 µg/ml

      Lane 1 : Untreated NIH/3T3 (mouse embryo), prepared in FBS free medium overnight, whole cell lysate
      Lane 2 : NIH/3T3, prepared in FBS free medium overnight, then treated with 10 µg/ml anisomycin for 30 minutes whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size : 54 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)


      Exposure time : 5 seconds

      Blocking and diluting buffer used was 2% BSA/TBST.

    • All lanes : Anti-ATF2 (phospho T71) antibody [E268] (ab32019) at 0.1 µg/ml

      Lane 1 : HeLa (human cervix adenocarcinoma), prepared in FBS free medium overnight, whole cell lysate
      Lane 2 : HeLa, prepared in FBS free medium overnight, then treated with 250ng/ml anisomycin for 30 minutes whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size : 54 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)


      Exposure time : 10 seconds

      Blocking and diluting buffer used was 2% BSA/TBST .
      The molecular weight is 70kD, consistent with the literature (PMID: 24223142).

    • Lane 1 (input): HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate  10μg.
      Lane 2 (+): HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.
      Lane 3 (-):  Rabbit monoclonal IgG (ab172730) instead of ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.

      Ab32019 Immunoprecipitating ATF2 in Human Hela whole cell lysate. For western blotting ab32019 (1:1000) was used to confirm successful immunoprecipitation. Blocking and diluting buffer used was 5% NFDM/TBST.

    References for Anti-ATF2 (phospho T71) antibody [E268] (ab32019)

    This product has been referenced in:

    See 1 Publication for this product

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