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RecombinantRabMAb

Recombinant Anti-ATF5 antibody [EPR18286] (ab184923)

Reviews (1) Submit a question References (10)
Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Flow Cytometry (Intracellular) - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Immunoprecipitation - Anti-ATF5 antibody [EPR18286] (ab184923)
  • Anti-ATF5 antibody [EPR18286] (ab184923)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18286] to ATF5
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 APC Carrier Free PE

Overview

  • Product name

    Anti-ATF5 antibody [EPR18286]
    See all ATF5 primary antibodies

  • Description

    Rabbit monoclonal [EPR18286] to ATF5

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 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.

Properties

Applications

The Abpromise guarantee

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)
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 (Intra)
Use at an assay dependent concentration.
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 (Intra)
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

    • Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)
      Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)
      Immunocytochemistry/ Immunofluorescence - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

    • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      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
      All lanes : 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 (Intracellular) - Anti-ATF5 antibody [EPR18286] (ab184923)
      Flow Cytometry (Intracellular) - Anti-ATF5 antibody [EPR18286] (ab184923)

      Intracellular 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. 

       

       

    • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      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
      All lanes : 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.

    • Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      Western blot - Anti-ATF5 antibody [EPR18286] (ab184923)
      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
      All lanes : 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

      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-ATF5 antibody [EPR18286] (ab184923)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

      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-ATF5 antibody [EPR18286] (ab184923)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

      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-ATF5 antibody [EPR18286] (ab184923)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF5 antibody [EPR18286] (ab184923)

      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.

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

    • Immunoprecipitation - Anti-ATF5 antibody [EPR18286] (ab184923)
      Immunoprecipitation - Anti-ATF5 antibody [EPR18286] (ab184923)

      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 Detection Reagent (HRP) (ab131366) was used for detection 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

    • Anti-ATF5 antibody [EPR18286] (ab184923)
      Anti-ATF5 antibody [EPR18286] (ab184923)

    References (10)

    Publishing research using ab184923? Please let us know so that we can cite the reference in this datasheet.

    ab184923 has been referenced in 10 publications.

    • Lee Y  et al. General anesthesia activates the mitochondrial unfolded protein response and induces age-dependent, long-lasting changes in mitochondrial function in the developing brain. Neurotoxicology 82:1-8 (2021). PubMed: 33144179
    • He F  et al. ATF5 and HIF1a cooperatively activate HIF1 signaling pathway in esophageal cancer. Cell Commun Signal 19:53 (2021). PubMed: 33980247
    • Jenkins EC  et al. Doxycycline promotes proteasome fitness in the central nervous system. Sci Rep 11:17003 (2021). PubMed: 34417525
    • Nandi SS  et al. Neurogenic Hypertension Mediated Mitochondrial Abnormality Leads to Cardiomyopathy: Contribution of UPRmt and Norepinephrine-miR- 18a-5p-HIF-1a Axis. Front Physiol 12:718982 (2021). PubMed: 34912235
    • Verma N  et al. The mRNA levels of heat shock factor 1 are regulated by thermogenic signals via the cAMP-dependent transcription factor ATF3. J Biol Chem 295:5984-5994 (2020). PubMed: 32184357
    • Kenny TC  et al. Mitohormesis Primes Tumor Invasion and Metastasis. Cell Rep 27:2292-2303.e6 (2019). PubMed: 31116976
    • Zhang X  et al. A novel role of endocan in alleviating LPS-induced acute lung injury. Life Sci 202:89-97 (2018). PubMed: 29627442
    • York D  et al. Expression and targeting of transcription factor ATF5 in dog gliomas. Vet Comp Oncol N/A:N/A (2017). Dog . 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 . PubMed: 28785242
    • Nukuda A  et al. Role of ATF5 in the invasive potential of diverse human cancer cell lines. Biochem Biophys Res Commun 474:509-514 (2016). PubMed: 27125458

    Customer reviews and Q&As

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    Western blot abreview for Anti-ATF5 antibody [EPR18286]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Nothobranchius furzeri Tissue lysate - whole (Liver and Skeletal muscle)
    Gel Running Conditions
    Non-reduced Denaturing (12% gel)
    Loading amount
    10 µg
    Specification
    Liver and Skeletal muscle
    Blocking step
    Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Jun 13 2019

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