Anti-PKA R2 (phospho S99) antibody [E151] (ab32390)

Overview

  • Product nameAnti-PKA R2 (phospho S99) antibody [E151]
    See all PKA R2 primary antibodies
  • Description
    Rabbit monoclonal [E151] to PKA R2 (phospho S99)
  • SpecificityThe antibody only detects PKA R2 phosphorylated on Serine 99. Phosphoserine 99 in Human corresponds to phosphoserine 96 in mouse.
  • Tested applicationsSuitable for: WB, IHC-P, IP, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PKA R2 (phospho S99).

  • Positive control
    • WB: K562 cell lysate. ICC/IF: HeLa cells. IHC-P: Human lympho node metastatic tissue.
  • 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 ab32390 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. Detects a band of approximately 51 kDa (predicted molecular weight: 45 kDa).
IHC-P 1/250.
IP 1/20.
ICC/IF 1/500.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionRegulatory subunit of the cAMP-dependent protein kinases involved in cAMP signaling in cells. Type II regulatory chains mediate membrane association by binding to anchoring proteins, including the MAP2 kinase.
    • Tissue specificityFour types of regulatory chains are found: I-alpha, I-beta, II-alpha, and II-beta. Their expression varies among tissues and is in some cases constitutive and in others inducible.
    • Sequence similaritiesBelongs to the cAMP-dependent kinase regulatory chain family.
      Contains 2 cyclic nucleotide-binding domains.
    • Post-translational
      modifications
      Phosphorylated by the activated catalytic chain.
    • Cellular localizationCytoplasm. Cell membrane. Colocalizes with PJA2 in the cytoplasm and the cell membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • cAMP dependent protein kinase regulatory subunit alpha 2 antibody
      • cAMP dependent protein kinase regulatory subunit RII alpha antibody
      • cAMP dependent protein kinase type II alpha regulatory chain antibody
      • cAMP dependent protein kinase type II alpha regulatory subunit antibody
      • cAMP-dependent protein kinase type II-alpha regulatory subunit antibody
      • KAP2 antibody
      • KAP2_HUMAN antibody
      • MGC3606 antibody
      • PKR 2 antibody
      • PKR2 antibody
      • PRKA R2 antibody
      • PRKAR 2 antibody
      • PRKAR2 antibody
      • PRKAR2A antibody
      • Protein kinase A RII alpha subunit antibody
      • Protein kinase cAMP dependent regulatory type II alpha antibody
      see all

    Anti-PKA R2 (phospho S99) antibody [E151] images

    • Cell line: HeLa (human cervix adenocarcinoma)

      Target AbID: Ab32390 anti-PKA R2 (phospho S99)

      Target Secondary: ab150077 AlexaFluor®488 goat anti-rabbit

      Counterstain AbID: ab7291 anti-Tubulin (mouse mAb)

      Target Secondary: ab150120 AlexaFluor®594 goat anti-mouse

      Fixative: 4% PFA

      Permeabilization: 0.1% Triton-X

      Nuclear counter stain: DAPI

      Other comments: Confocal image showed increased cytoplasmic staining after CA (100ng/ml, 10min) treatment on Hela cells. The LP treatment decreased the increased cytoplasmic staining caused by CA.  Ab32514 was used as a Pan control for ab32390. The results showed cytoplasmic staining on untreated, CA and CA+LP treated Hela cells. 

    • All lanes : Anti-PKA R2 (phospho S99) antibody [E151] (ab32390) at 1/2000 dilution

      Lane 1 : K562 whole cell lysate with PKA R2 (pS99) phospho peptide
      Lane 2 : K562 whole cell lysate with PKA R2 non-phospho peptide

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

      Predicted band size : 45 kDa
      Observed band size : 51 kDa (why is the actual band size different from the predicted?)


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-PKA R2 (phospho S99) antibody [E151] (ab32390) at 1/10000 dilution

      Lane 1 : Untreated K562 whole cell lysate
      Lane 2 : K562 whole cell lysate - treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 45 kDa
      Observed band size : 51 kDa (why is the actual band size different from the predicted?)


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • ab32390, at a 1/250 dilution, staining (A) untreated (B) Phosphatase treated lymph node metastatic by immunohistochemistry, Paraffin embedded tissue

    References for Anti-PKA R2 (phospho S99) antibody [E151] (ab32390)

    This product has been referenced in:
    • Isensee J  et al. Pain modulators regulate the dynamics of PKA-RII phosphorylation in subgroups of sensory neurons. J Cell Sci 127:216-29 (2014). Read more (PubMed: 24190886) »
    • Bantubungi K  et al. Cdkn2a/p16Ink4a regulates fasting-induced hepatic gluconeogenesis through the PKA-CREB-PGC1a pathway. Diabetes 63:3199-209 (2014). Read more (PubMed: 24789920) »

    See all 7 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (vero)
    Loading amount 20000 cells
    Specification vero
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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    Verified customer

    Submitted Jun 23 2009

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