Overview

  • Product name

    Anti-ATF2 antibody [EPR22938-114]
    See all ATF2 primary antibodies
  • Description

    Rabbit monoclonal [EPR22938-114] to ATF2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IP, IHC-Pmore details
    Unsuitable for: ChIP or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse ATF2 aa 100-350. The exact sequence is proprietary.
    Database link: P16951

  • Positive control

    • WB: NIH/3T3, RAW 264.7, HEK-293T, K562, PC-12 and Jurkat whole cell lysate. Mouse brain tissue lysate. Rat spleen tissue lysate. IHC-P: Human kidney carcinoma tissue. Human, mouse and rat kidney tissue. Flow Cyt: RAW 264.7 and NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
  • General notes

    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 ab239361 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 1/40.
WB 1/1000.
IP 1/30.
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ChIP or ICC/IF.
  • Target

    • Function

      Transcriptional activator, probably constitutive, which binds to the cAMP-responsive element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Interaction with JUN redirects JUN to bind to CRES preferentially over the 12-O-tetradecanoylphorbol-13-acetate response elements (TRES) as part of an ATF2/JUN complex.
    • Tissue specificity

      Abundant expression seen in the brain.
    • Sequence similarities

      Belongs to the bZIP family. ATF subfamily.
      Contains 1 bZIP domain.
      Contains 1 C2H2-type zinc finger.
    • Post-translational
      modifications

      Phosphorylation of Thr-69 and Thr-71 by MAPK14 causes increased transcriptional activity. Also phosphorylated and activated by JNK.
    • Cellular localization

      Nucleus.
    • Information by UniProt
    • Database links

    • Alternative names

      • Activating transcription factor 2 antibody
      • Activating transcription factor 2 splice variant ATF2 var2 antibody
      • ATF 2 antibody
      • Atf-2 antibody
      • Atf2 antibody
      • ATF2 protein antibody
      • ATF2_HUMAN antibody
      • cAMP Response Element Binding Protein 2 antibody
      • cAMP response element binding protein CRE BP1 antibody
      • cAMP response element-binding protein CRE-BP1 antibody
      • cAMP responsive element binding protein 2, formerly antibody
      • cAMP-dependent transcription factor ATF-2 antibody
      • cAMP-responsive element-binding protein 2 antibody
      • CRE BP1 antibody
      • CRE-BP antibody
      • CREB 2 antibody
      • CREB-2 antibody
      • CREB2 antibody
      • CREBP1 antibody
      • Cyclic AMP dependent transcription factor ATF 2 antibody
      • Cyclic AMP-dependent transcription factor ATF-2 antibody
      • Cyclic AMP-responsive element-binding protein 2 antibody
      • D130078H02Rik antibody
      • D18875 antibody
      • HB 16 antibody
      • HB16 antibody
      • Histone acetyltransferase ATF2 antibody
      • MGC105211 antibody
      • MGC105222 antibody
      • MGC111558 antibody
      • MGC142504 antibody
      • mXBP antibody
      • MXBP protein antibody
      • Tg(Gzma-Klra1)7Wum antibody
      • TREB 7 antibody
      • TREB7 antibody
      see all

    Images

    • All lanes : Anti-ATF2 antibody [EPR22938-114] (ab239361) at 1/1000 dilution

      Lane 1 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
      Lane 2 : Rat spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Observed band size: 70 kDa
      why is the actual band size different from the predicted?



      The molecular weight and degraded fragments observed are consistent with what has been described in the literature (PMID: 9488727, 10207054, 26901653).  Suggest use freshly made lysate to minimize protein degradation.

      Exposure time: 3 minutes.

      Blocking and Diluting Buffer and concentration: 5% NFDM/TBST.

    • Anti-ATF2 antibody [EPR22938-114] (ab239361) at 1/1000 dilution + Mouse brain tissue lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Observed band size: 70 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID:26901653).

      Exposure time: 3 minutes.

      Blocking and Diluting Buffer and concentration: 5% NFDM/TBST.

    • All lanes : Anti-ATF2 antibody [EPR22938-114] (ab239361) at 1/1000 dilution

      Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
      Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
      Lane 3 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
      Lane 4 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
      Lane 5 : Jurkat (human T cell leukemia T lymphocyte), 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

      Observed band size: 70 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID:26901653).

      Exposure time: 10 seconds.

      Blocking and Diluting Buffer and concentration: 5% NFDM/TBST.

    • ATF2 was immunoprecipitated from 0.35 mg NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate with ab239361 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239361. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

      Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10µg.

      Lane 2: ab239361 IP in NIH/3T3 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239361 in NIH/3T3 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 8 seconds

      Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

      The molecular weight and degraded fragments observed are consistent with what has been described in the literature (PMID: 9488727, PMID: 10207054, PMID:26901653).

    • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling ATF2 with ab239361 at 1/40 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling ATF2 with ab239361 at 1/40 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling ATF2 with ab239361 at 1/250 dilution (1.76 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat kidney is observed. The section was incubated with ab239361 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ATF2 with ab239361 at 1/250 dilution (1.76 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse kidney is observed. The section was incubated with ab239361 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemical analysis of paraffin-embedded human kidney carcinoma tissue labeling ATF2 with ab239361 at 1/250 dilution (1.76 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human kidney carcinoma (PMID: 27377902) is observed. The section was incubated with ab239361 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ATF2 with ab239361 at 1/250 dilution (1.76 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human kidney (PMID: 27377902) is observed. The section was incubated with ab239361 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    References

    ab239361 has not yet been referenced specifically in any publications.

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