Recombinant
RabMAb

Recombinant Anti-ATF-4 antibody [EPR18111] (ab184909)

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Overview

  • Product name

    Anti-ATF-4 antibody [EPR18111]
    See all ATF-4 primary antibodies

  • Description

    Rabbit monoclonal [EPR18111] to ATF-4

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat

  • Immunogen

    Recombinant fragment within Human ATF-4 aa 1-200. The exact sequence is proprietary.
    Database link: P18848

  • Positive control

    • WB: Human fetal liver and fetal kidney lysates; A431, HeLa, whole cell lysates; untransfected HEK-293 whole cell lysate (control) lysate; HEK-293 whole cell lysate transfected with ATF-4 with GFP-tag lysate; HepG2 treated with 5 µM MG-132 (ab141003) and 3 µg/ml tunicamycin (ab120296) for 6 hours whole cell lysate. IHC-P: Human bladder cancer and stomach cancer tissues. ICC/IF: HeLa and A431 cells. Flow Cyt: HeLa cells. IP: HeLa 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.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR18111

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab184909 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: 38 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/80.
IP 1/50.

Target

  • Function

    Transcriptional activator. Binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. It binds to a Tax-responsive enhancer element in the long terminal repeat of HTLV-I.

  • Sequence similarities

    Belongs to the bZIP family.
    Contains 1 bZIP domain.

  • Cellular localization

    Cytoplasm. Cell membrane. Nucleus. Colocalizes with GABBR1 in hippocampal neuron dendritic membranes.
  • Information by UniProt

  • Database links

    • Alternative names

      • Activating transcription factor 4 antibody
      • ATF 4 antibody
      • ATF4 antibody
      • ATF4 protein antibody
      • ATF4_HUMAN antibody
      • cAMP-dependent transcription factor ATF-4 antibody
      • cAMP-responsive element-binding protein 2 antibody
      • CREB 2 antibody
      • CREB-2 antibody
      • CREB2 antibody
      • Cyclic AMP dependent transcription factor ATF 4 antibody
      • Cyclic AMP response element binding protein 2 antibody
      • Cyclic AMP-dependent transcription factor ATF-4 antibody
      • Cyclic AMP-responsive element-binding protein 2 antibody
      • DNA binding protein TAXREB67 antibody
      • DNA-binding protein TAXREB67 antibody
      • Tax Responsive Enhancer Element B67 antibody
      • Tax-responsive enhancer element-binding protein 67 antibody
      • TaxREB67 antibody
      • TXREB antibody
      see all

    Images

    • Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/1000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
      Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 4 : Human fetal kidney lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

      Predicted band size: 38 kDa
      Observed band size: 50 kDa
      why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 3 minutes; Lane 2 and 3: 30 seconds; Lane 4: 3 minutes.

    • Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATF-4 with ab184909 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear staining on HeLa cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab184909 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

      -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)This image was courtesy of an annoymous AbReview

      Formalin-fixed paraffin-embedded human colon cancer tissue stained for ATF-4 using ab184909 at 1/80 dilution (45 minute incubation) in immunohistochemical analysis. A polyclonal Goat Horse Radish Peroxidase antibody was used as the secondary.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)

      Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue labeling ATF-4 with ab184909 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

      Nucleus staining on human stomach cancer is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/5000 dilution

      Lane 1 : Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control)
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with ATT-4 with GFP-tag 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: 38 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
      All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/1000 dilution

      Lane 1 : Untreated HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (control)
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) treated with 5 µM MG-132 (ab141003) and 3 µg/ml tunicamycin (ab120296) for 6 hours 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: 38 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with what has been described in the literature (PMID: 22095285).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)

      Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling ATF-4 with ab184909 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

      Nucleus staining on human bladder cancer is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling ATF-4 with ab184909 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear staining on A431 cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab184909 at 1/1000 dilution followed by ab150120  at 1/1000 dilution.

      -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    • Flow Cytometry - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Flow Cytometry - Anti-ATF-4 antibody [EPR18111] (ab184909)

      Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATF-4 with ab184909 at 1/80 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-ATF-4 antibody [EPR18111] (ab184909)
      Immunoprecipitation - Anti-ATF-4 antibody [EPR18111] (ab184909)

      ATF-4 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184909 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab184909 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HeLa whole cell lysate, 10µg (Input).

      Lane 2: ab184909 IP in HeLa whole cell lysate.

      Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184909 in HeLa whole cell lysate

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

      Exposure time: 3 minutes.

    References

    This product has been referenced in:

    • Li Z  et al. Lipofectamine 2000/siRNA complexes cause endoplasmic reticulum unfolded protein response in human endothelial cells. J Cell Physiol N/A:N/A (2019). Read more (PubMed: 31032939) »
    • Wang MG  et al. Activation of PERK-eIF2a-ATF4-CHOP axis triggered by excessive ER stress contributes to lead-induced nephrotoxicity. Biochim Biophys Acta Mol Cell Res N/A:N/A (2018). Read more (PubMed: 30528975) »
    See all 10 Publications for this product

    Publishing research using ab184909? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATF-4 antibody [EPR18111]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Colon cancer)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
    Permeabilization
    No
    Specification
    Colon cancer
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 19 2018

    Western blot abreview for Anti-ATF-4 antibody [EPR18111]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (OSW cells)
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Loading amount
    100 µg
    Specification
    OSW cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Ms. Liubov Lobanova

    Verified customer

    Submitted Jul 22 2016

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