Overview

  • Product name
    Anti-ATF5 antibody [EPR18286]
    See all ATF5 primary antibodies
  • Description
    Rabbit monoclonal [EPR18286] to ATF5
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ATF5 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9Y2D1

  • Positive control
    • WB: Jurkat, HEK-293, SH-SY5Y, Neuro-2a, C6, PC-12, RAW 264.7 and NIH/3T3 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; Rat and mouse brain, heart and kidney lysates. IHC-P: Human breast, Human hepatocellular carcinoma, mouse cardiac muscle and rat stomach tissues. ICC/IF: NIH/3T3 and Jurkat cells. IP: NIH/3T3 whole cell lysate
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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

Applications

Our Abpromise guarantee covers the use of ab184923 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 1/2000. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
IP 1/50.
Flow Cyt Use at an assay dependent concentration.

Target

  • Relevance
    ATF5 or Activating transcription factor 5, binds to cAMP inducible promoters and is involved in gene transcription. This protein binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. ATF5 plays a role in inhibition of nerve growth factor induced neuronal outgrowth and regulation of neurogenesis.
  • Cellular localization
    Cytoplasmic and Nuclear
  • Database links
  • Alternative names
    • Activating transcription factor 5 alpha/beta antibody
    • Activating transcription factor 5 antibody
    • Activating transcription factor X antibody
    • AFTA antibody
    • ATF 5 antibody
    • ATF 7 antibody
    • ATF7 antibody
    • ATFX antibody
    • BZIP protein ATF7 antibody
    • cAMP dependent transcription factor ATF 5 antibody
    • cAMP dependent transcription factor ATF5 antibody
    • Cyclic AMP dependent transcription factor ATF 5 antibody
    • Cyclic AMP dependent transcription factor ATF5 antibody
    • FLJ34666 antibody
    • HMFN0395 antibody
    • MGC102397 antibody
    • NAP1 antibody
    • NRIF3 associated protein antibody
    • ODA 10 antibody
    • Transcription factor ATFx antibody
    • Transcription factor like protein ODA 10 antibody
    • Transcription factor like protein ODA10 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling ATF5 with ab184923 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on Jurkat cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab184923 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling ATF5 with ab184923 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab184923 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • All lanes : Anti-ATF5 antibody [EPR18286] (ab184923) at 1/2000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 3 : SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate
    Lane 4 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 31 kDa
    Observed band size : 31 kDa


    Exposure time : 10 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling ATF5 with purified ab184923 at 1/120 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • All lanes : Anti-ATF5 antibody [EPR18286] (ab184923) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size : 31 kDa
    Observed band size : 31 kDa


    Exposure time : 10 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-ATF5 antibody [EPR18286] (ab184923) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab136636) at 1/1000 dilution

    Predicted band size : 31 kDa
    Observed band size : 31 kDa


    Exposure time : 10 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ATF5 with ab184923 at 1/1000 dilution, followed  by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and weak cytoplasm staining on Human breast 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.

  • Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling ATF5 with ab184923 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and weak cytoplasm staining on tumor cells of hepatocellular carcinoma 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.

  • Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling ATF5 with ab184923 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on mouse cardiac muscle 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.

  • Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling ATF5 with ab184923 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on rat stomach 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.

  • ATF5 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab184923 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab184923 at 1/10000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1500 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 µg (Input). Lane 2: ab184923 IP in NIH/3T3 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184923 in NIH/3T3 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second

References

This product has been referenced in:
  • York D  et al. Expression and targeting of transcription factor ATF5 in dog gliomas. Vet Comp Oncol N/A:N/A (2017). Dog . Read more (PubMed: 28480569) »
  • Ben-Shmuel S  et al. Activating Transcription Factor-5 Knockdown Reduces Aggressiveness of Mammary Tumor Cells and Attenuates Mammary Tumor Growth. Front Endocrinol (Lausanne) 8:173 (2017). WB ; Mouse . Read more (PubMed: 28785242) »

See all 2 Publications for this product

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