RecombinantRabMAb

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

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18111] to ATF-4
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

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

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

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.

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

    References (26)

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

    ab184909 has been referenced in 26 publications.

    • Arrillaga-Romany I  et al. Biological activity of weekly ONC201 in adult recurrent glioblastoma patients. Neuro Oncol 22:94-102 (2020). PubMed: 31702782
    • Tian F  et al. KLF6 Induces Apoptosis in Human Lens Epithelial Cells Through the ATF4-ATF3-CHOP Axis. Drug Des Devel Ther 14:1041-1055 (2020). PubMed: 32210535
    • Shen T  et al. Pancreatic cancer-derived exosomes induce apoptosis of T lymphocytes through the p38 MAPK-mediated endoplasmic reticulum stress. FASEB J 34:8442-8458 (2020). PubMed: 32350913
    • Fang K  et al. Enhanced expression of asparagine synthetase under glucose-deprived conditions promotes esophageal squamous cell carcinoma development. Int J Med Sci 17:510-516 (2020). PubMed: 32174781
    • Zhao Y  et al. AQP1 suppression by ATF4 triggers trabecular meshwork tissue remodelling in ET-1-induced POAG. J Cell Mol Med 24:3469-3480 (2020). PubMed: 32052937
    View all Publications for this product

    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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