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

Overview

  • Product nameAnti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y]
  • Description
    Rabbit monoclonal [EP2606Y] to PKA alpha/beta/gamma (catalytic subunit) (phospho T197)
  • SpecificityDetects PKA alpha/beta/gamma (catalytic subunit) phosphorylated on threonine 197. Moreover, the antibody will cross-react with PRKX/Y (pT203).
  • Tested applicationsSuitable for: WB, IP, IHC-P, ICCmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control
    • WB: HeLa cell lysate treated with calyculin A.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

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.

Application Abreviews Notes
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.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

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

    • 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
      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.

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      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.

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

    This product has been referenced in:
    • Ferdaus MZ  et al. SPAK and OSR1 play essential roles in potassium homeostasis through actions on the distal convoluted tubule. J Physiol 594:4945-66 (2016). Read more (PubMed: 27068441) »
    • Han YS  et al. Human heart failure is accompanied by altered protein kinase A subunit expression and post-translational state. Arch Biochem Biophys 538:25-33 (2013). Read more (PubMed: 23942052) »

    See all 3 Publications for this product

    Product Wall

    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
    Username

    Dr. Karine Thibault

    Verified customer

    Submitted Nov 05 2014

    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 th...

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