Overview

  • Product name

    Anti-PKA R2/PKR2 (phospho S99) antibody [E151]
    See all PKA R2/PKR2 primary antibodies
  • Description

    Rabbit monoclonal [E151] to PKA R2/PKR2 (phospho S99)
  • Host species

    Rabbit
  • Specificity

    The antibody only detects PKA R2/PKR2 phosphorylated on Serine 99. Phosphoserine 99 in Human corresponds to phosphoserine 96 in mouse.

    The mouse and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse and pig.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    corresponding to Human PKA R2/PKR2 (phospho S99).

  • Positive control

    • WB: K562 cell lysate. HeLa starved for 3 hours and then treated with 100 nM Calyculin A for 30 minutes, NIH/3T3, NIH/3T3 treated with Calyculin A, and C6 treated with 100 ng/ml Calyculin A for 1 hour cell lysates. Pig heart lysates. ICC/IF: HeLa cells. IHC-P: Human lymph node metastatic tissue. Human kidney without Lambda Protein Phosphatase treatment (image A) and Rat kidney without Lambda Protein Phosphatase treatment
  • General notes

     

     

     This product was previously labelled as PKA R2

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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 1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 45 kDa).

For unpurified use at 1/2500

IHC-P 1/500.

The mouse and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse and pig.

See IHC antigen retrieval protocols.

For unpurified use at 1/250

ICC/IF 1/500.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function

      Regulatory 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 specificity

      Four 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 similarities

      Belongs to the cAMP-dependent kinase regulatory chain family.
      Contains 2 cyclic nucleotide-binding domains.
    • Post-translational
      modifications

      Phosphorylated by the activated catalytic chain.
    • Cellular localization

      Cytoplasm. 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

    Images

    • All lanes : Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390) at 1/1000 dilution (Purified)

      Lane 1 : Pig heart lysates
      Lane 2 : Pig heart. 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: 45 kDa
      Observed band size: 51 kDa
      why is the actual band size different from the predicted?

    • All lanes : Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390) at 1/1000 dilution (Purified)

      Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
      Lane 2 : C6 (Rat glial tumor glial cell) treated with 100ng/ml Calyculin A for 1 hour
      Lane 3 : C6 (Rat glial tumor glial cell) treated with 100ng/ml Calyculin A for 1 hour.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: 45 kDa
      Observed band size: 51 kDa why is the actual band size different from the predicted?

    • All lanes : Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390) at 1/1000 dilution (Purified)

      Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with Calyculin A
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with Calyculin A. 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: 45 kDa
      Observed band size: 51 kDa why is the actual band size different from the predicted?

    • All lanes : Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) were starved for 3 hours and then treated with 100 nM Calyculin A for 30 minutes
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100 nM Calyculin A for 30 minutes. 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: 45 kDa
      Observed band size: 51 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney without Lambda Protein Phosphatase treatment (image A) and treated with Lambda Protein Phosphatase (image B). tissue sections labeling PKA R2/PKR2 with purified ab32390 at 1:500 dilution (0.75 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney without Lambda Protein Phosphatase treatment (image A) and treated with Lambda Protein Phosphatase (image B). tissue sections labeling PKA R2/PKR2 with purified ab32390 at 1:500 dilution (0.75 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • All lanes : Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390) at 1/2000 dilution

      Lane 1 : K562 whole cell lysate with PKA R2/PKR2 (pS99) phospho peptide
      Lane 2 : K562 whole cell lysate with PKA R2/PKR2 non-phospho peptide

      Secondary
      All lanes : 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.

    • Cell line: HeLa (human cervix adenocarcinoma)

      Target AbID: Ab32390 anti-PKA R2/PKR2 (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/PKR2 (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
      All lanes : 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

    This product has been referenced in:

    • Speichert S  et al. Role of Norepinephrine in IL-1ß-Induced Chondrocyte Dedifferentiation under Physioxia. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30861996) »
    • Isensee J & Hucho T High-Content Imaging of Immunofluorescently Labeled TRPV1-Positive Sensory Neurons. Methods Mol Biol 1987:111-124 (2019). Read more (PubMed: 31028677) »
    See all 16 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (vero)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20000 cells
    Specification
    vero
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Jun 23 2009

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up