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RabMAb

Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) 抗体 [EP2606Y] (ab75991)

製品の概要

  • 製品名
    Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y]
    PKA alpha/beta/gamma (catalytic subunit) 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP2606Y] to PKA alpha/beta/gamma (catalytic subunit) (phospho T197)
  • 由来種
    Rabbit
  • 特異性
    Detects PKA alpha/beta/gamma (catalytic subunit) phosphorylated on threonine 197. Moreover, the antibody will cross-react with PRKX/Y (pT203).
  • アプリケーション
    適用あり: WB, IP, IHC-P, ICC, Dot blotmore details
    適用なし: Flow Cyt
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human PKA alpha/beta/gamma (catalytic subunit) aa 150-250 (phospho T197). The exact sequence is proprietary.

  • ポジティブ・コントロール
    • WB: HeLa cell lysate treated with calyculin A.
  • 特記事項

     

     

    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.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab75991 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/2500 - 1/5000. Predicted molecular weight: 40 kDa.
IP 1/30.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use of HRP-conjugated or polymerized HRP secondary antibodies recommended, stronger signals have been found using the polymerized HRP secondary.

ICC 1/100 - 1/250.
Dot blot Use at an assay dependent concentration.
  • 追加情報
    Is unsuitable for Flow Cyt.
  • ターゲット情報

    画像

    • All lanes : Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (ab75991) at 1/1000 dilution

      Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) 10% FBS all the time. Whole cell lysates
      Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) serum starve for 24h then 20% FBS 20h. Whole cell lysates
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) serum starve for 24h then 20% FBS 20h.Whole cell lysates.Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 40 kDa
      Observed band size: 40 kDa


      Exposure time: 1 second


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

    • Dot blot analysis of various peptides labelling PKA gamma (catalytic subunit) (phospho T197) with ab75991 at a dilution of 1/1000. ab97051, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/100000).

      Lane 1 - PKA alpha/beta/gamma (catalytic subunit) (pT197) phospho peptide.

      Lane 2 - PKA alpha/beta/gamma (catalytic subunit) non-phospho peptide.

      Lane 3 - PRKX/Y (pT203) phospho peptide.

      Lane 4 - PRKX/Y non-phospho peptide.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • All lanes : Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (ab75991) at 1/5000 dilution

      Lane 1 : HeLa whole cell lysate - treated with Calyculin A with PKA alpha/beta/gamma (catalytic subunit) (pT197) phospho peptide
      Lane 2 : HeLa whole cell lysate - treated with Calyculin A with PKA alpha/beta/gamma (catalytic subunit) non-phospho peptide
      Lane 3 : HeLa whole cell lysate - treated with Calyculin A with PRKX/Y (pT203) phospho peptide
      Lane 4 : HeLa whole cell lysate - treated with Calyculin A with PRKX/Y non- phospho peptide

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 40 kDa
      Observed band size: 40 kDa


      Exposure time: 3 minutes


      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (ab75991) at 1/5000 dilution

      Lane 1 : HeLa cell lysates, untreated
      Lane 2 : HeLa cell lysates treated with calyculin A

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat-anti rabbit IgG at 1/2000 dilution

      Predicted band size: 40 kDa
      Observed band size: 40 kDa

    • ab75991, at a 1/100 dilution, staining PKA gamma (catalytic subunit) in formalin fixed, paraffin embedded human testis tissue by Immunohistochemistry.

    参考文献

    This product has been referenced in:
    • Ding Q  et al. Lack of endogenous parathyroid hormone delays fracture healing by inhibiting vascular endothelial growth factor-mediated angiogenesis. Int J Mol Med 42:171-181 (2018). Read more (PubMed: 29620150) »
    • Cao N  et al. Clinical-grade human umbilical cord-derived mesenchymal stem cells reverse cognitive aging via improving synaptic plasticity and endogenous neurogenesis. Cell Death Dis 8:e2996 (2017). Read more (PubMed: 28796260) »
    See all 11 Publications for this product

    レビューと Q&A

    1-2 of 2 Abreviews or Q&A

    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (gel 10% SDS-PAGE)
    Sample
    Mouse Tissue lysate - whole (Brain)
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Dr. Karine Thibault

    Verified customer

    投稿 Nov 05 2014

    Answer

    Thank you for contacting us.

    In GPCR signalling pathway cAMP binds to the regulatory subunit of PKA (Protein Kinase-A) leading to the dissociation of the catalytic subunits thereby making it active. PKA then lead to the phosphorylation of the GPCR for further cellular signalling.

    Once phosphorylated PKA become active, this antibody detects the phophorylated form of PKA hence the active catalytic subunit of PKA.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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