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    atf-4-antibody-epr18111-ab184909.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper Leucine Zipper
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RecombinantRabMAb

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

  • Datasheet
  • Certificate of Compliance
Reviews (4) Submit a question References (44)

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Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • 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)
  • Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Flow Cytometry (Intracellular) - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Immunoprecipitation - Anti-ATF-4 antibody [EPR18111] (ab184909)
  • Anti-ATF-4 antibody [EPR18111] (ab184909)

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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

APC Carrier Free FITC PE

You may also be interested in

Protein
Product image
Recombinant Human ATF-4 protein (ab109946)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
Product image
Anti-ATF-4 antibody [EPR18111] - BSA and Azide free (ab221791)

View more associated products

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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

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

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra): 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.

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

    pH: 7.2
    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

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • Leucine Zipper
    • Signal Transduction
    • Antibodies
    • atf-4

Associated products

  • Alternative Versions

    • Anti-ATF-4 antibody [EPR18111] - BSA and Azide free (ab221791)
    • APC Anti-ATF-4 antibody [EPR18111] (ab225331)
    • FITC Anti-ATF-4 antibody [EPR18111] (ab225332)
    • PE Anti-ATF-4 antibody [EPR18111] (ab225333)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human ATF-4 protein (ab109946)
  • Related Buffer

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Tunicamycin, Protein glycosylation inhibitor (ab120296)
    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • MG-132, proteasome inhibitor (ab141003)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/80.
WB (2)
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 38 kDa).
IHC-P (1)
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/50.
Notes
Flow Cyt (Intra)
1/80.
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.
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.
  • Target information above from: UniProt accession P18848 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 468 Human
    • Omim: 604064 Human
    • SwissProt: P18848 Human
    • Unigene: 496487 Human
    • 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.

      See Abreview

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

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell linefrom cervix adenocarcinoma) cells labeling ATF-4with 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)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (44)

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

    ab184909 has been referenced in 44 publications.

    • Arioka Y  et al. Chromosome 22q11.2 deletion causes PERK-dependent vulnerability in dopaminergic neurons. EBioMedicine 63:103138 (2021). PubMed: 33341442
    • Li KK  et al. U three protein 14a (UTP14A) promotes tumour proliferation and metastasis via the PERK/eIF2a/GRP78 signalling pathway in oesophageal squamous cell carcinoma. J Cancer 12:134-140 (2021). PubMed: 33391409
    • Oku Y  et al. Homeostasis of the ER redox state subsequent to proteasome inhibition. Sci Rep 11:8655 (2021). PubMed: 33883613
    • Bao J  et al. Pharmacological Disruption of Phosphorylated Eukaryotic Initiation Factor-2a/Activating Transcription Factor 4/Indian Hedgehog Protects Intervertebral Disc Degeneration via Reducing the Reactive Oxygen Species and Apoptosis of Nucleus Pulposus Cells. Front Cell Dev Biol 9:675486 (2021). PubMed: 34164397
    • Rong K  et al. Alendronate Alleviated Femoral Head Necrosis and Upregulated BMP2/EIF2AK3/EIF2A/ATF4 Pathway in Liquid Nitrogen Treated Rats. Drug Des Devel Ther 15:1717-1724 (2021). PubMed: 33935494
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    50 µg
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2021

    ChIP abreview for Anti-ATF-4 antibody [EPR18111]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (HEK293)
    Negative control
    No antibody
    Specification
    HEK293
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1.1% Formaldehyde
    Positive control
    Histone H3 antibody
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2021

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

    Excellent
    Abreviews
    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
    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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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