Overview

  • Product name

    Anti-ULK1 (phospho S757) antibody [EPR22265-9]
    See all ULK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22265-9] to ULK1 (phospho S757)
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse
  • Immunogen

    Synthetic peptide within Mouse ULK1 aa 750-850. The exact sequence is proprietary.
    Database link: O70405

  • Positive control

    • WB: MEF whole cell lysate. IP: MEF 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 ab229909 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 1/1000.
Dot blot 1/1000.
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Target

    • Function

      Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2 and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. May also phosphorylate SESN2 and SQSTM1 to regulate autophagy (PubMed:25040165).
    • Tissue specificity

      Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung.
    • Sequence similarities

      Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      Autophosphorylated. Phosphorylated under nutrient-rich conditions; dephosphorylated during starvation or following treatment with rapamycin. Under nutrient sufficiency, phosphorylated by MTOR/mTOR, disrupting the interaction with AMPK and preventing activation of ULK1 (By similarity). In response to nutrient limitation, phosphorylated and activated by AMPK, leading to activate autophagy.
    • Cellular localization

      Cytoplasm, cytosol. Preautophagosomal structure. Under starvation conditions, is localized to puncate structures primarily representing the isolation membrane that sequesters a portion of the cytoplasm resulting in the formation of an autophagosome.
    • Information by UniProt
    • Database links

    • Alternative names

      • ATG 1 antibody
      • ATG1 antibody
      • ATG1 autophagy related 1 homolog antibody
      • ATG1A antibody
      • Autophagy related protein 1 homolog antibody
      • Autophagy-related protein 1 homolog antibody
      • FLJ38455 antibody
      • FLJ46475 antibody
      • hATG1 antibody
      • KIAA0722 antibody
      • Serine/threonine protein kinase ULK1 antibody
      • Serine/threonine protein kinase Unc51.1 antibody
      • Serine/threonine-protein kinase ULK1 antibody
      • ULK 1 antibody
      • ULK1 antibody
      • ULK1_HUMAN antibody
      • Unc 51 (C. elegans) like kinase 1 antibody
      • UNC 51 antibody
      • Unc 51 like kinase 1 antibody
      • Unc-51 like kinase 1 (C. elegans) antibody
      • Unc-51-like kinase 1 antibody
      • UNC51 antibody
      • UNC51, C. elegans, homolog of antibody
      • Unc51.1 antibody
      see all

    Images

    • All lanes : Anti-ULK1 (phospho S757) antibody [EPR22265-9] (ab229909) at 1/1000 dilution

      Lane 1 : MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate
      Lane 2 : MEF whole cell lysate (Phosphatase treated membrane)

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



      WB membrane in lane 2 was treated with alkaline phosphatase for 1h at 37°C.  This blot was developed using a higher sensitivity ECL substrate.

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

      Exposure time: 59 seconds.

    • All lanes : Anti-ULK1 (phospho S757) antibody [EPR22265-9] (ab229909) at 1/1000 dilution

      Lane 1 : Untreated MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate
      Lane 2 : MEF cells were cultured in amino acid and serum-free, glucose-containing starvation buffer (EBSS) for 16 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

      Observed band size: 150 kDa why is the actual band size different from the predicted?


      Exposure time: 59 seconds


      Phosphorylation level of S757 is reduced during starvation (PMID: 28835610, 21258367). This blot was developed using a higher sensitivity ECL substrate.

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

      Exposure time: 59 seconds.

    • Dot blot analysis using ab229909 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 diltution.

      Lane 1: ULK1 (phospho S757) peptide.

      Lane 2: ULK1 non-phospho peptide.

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

      Exposure time: 3 minutes.

    • ULK1 (phospho S757) was immunoprecipitated from 0.35 mg MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate with ab229909 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab229909 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution. Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

      Lane 1: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate, 10µg.

      Lane 2: ab229909 IP in MEF whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229909 in MEF whole cell lysate.

      Blocking/dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 mins.

       

    References

    ab229909 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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