Recombinant
RabMAb

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

Overview

  • Product name

    Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y]
    See all PKA alpha/beta/gamma (catalytic subunit) primary antibodies
  • Description

    Rabbit monoclonal [EP2606Y] to PKA alpha/beta/gamma (catalytic subunit) (phospho T197)
  • Host species

    Rabbit
  • Specificity

    Detects PKA alpha/beta/gamma (catalytic subunit) phosphorylated on threonine 197. Moreover, the antibody will cross-react with PRKX/Y (pT203).
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC, Dot blotmore 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

     

     

    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 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.
Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    Images

    • 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.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Zhao H  et al. Single-Cell Transcriptomics of Human Oocytes: Environment-Driven Metabolic Competition and Compensatory Mechanisms During Oocyte Maturation. Antioxid Redox Signal 30:542-559 (2019). Read more (PubMed: 29486586) »
    • Xue B  et al. Astragaloside IV Protects Primary Cerebral Cortical Neurons from Oxygen and Glucose Deprivation/Reoxygenation by Activating the PKA/CREB Pathway. Neuroscience 404:326-337 (2019). Read more (PubMed: 30708047) »
    See all 20 Publications for this product

    Customer reviews and Q&As

    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

    Submitted 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.

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