Overview

  • Product name

    Anti-ATF-4 antibody [EPR18111] - BSA and Azide free
    See all ATF-4 primary antibodies
  • Description

    Rabbit monoclonal [EPR18111] to ATF-4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant fragment 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 MG132 and 3 µg/ml tunicamycin 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

    Ab221791 is the carrier-free version of ab184909. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab221791 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Applications

Our Abpromise guarantee covers the use of ab221791 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 Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 38 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184909).

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184909).

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184909).

References

ab221791 has not yet been referenced specifically in any publications.

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