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RecombinantRabMAb

Recombinant Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

Reviews (1) Submit a question References (30)
Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
  • Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E151] to PKA R2/PKR2 (phospho S99)
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human, Pig

Conjugates logo Related conjugates and formulations

Carrier Free

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

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

Applications

The Abpromise guarantee

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

For unpurified use at 1/2500
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    see all

  • 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

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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?

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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?

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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?

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

    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) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

    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.

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Immunocytochemistry/ Immunofluorescence - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

    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. 

  • Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Western blot - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

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

  • Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)
    Anti-PKA R2/PKR2 (phospho S99) antibody [E151] (ab32390)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (30)

Publishing research using ab32390? Please let us know so that we can cite the reference in this datasheet.

ab32390 has been referenced in 30 publications.

  • Li T  et al. Sevoflurane protects against ischemia-reperfusion injury in mice after total knee arthroplasty via facilitating RASD1-mediated protein kinase A pathway activation. Aging (Albany NY) 13:13333-13348 (2021). PubMed: 33982674
  • Garza Carbajal A  et al. Oncostatin M induces hyperalgesic priming and amplifies signaling of cAMP to ERK by RapGEF2 and PKA. J Neurochem 157:1821-1837 (2021). PubMed: 32885411
  • Hedderich J  et al. Norepinephrine Inhibits the Proliferation of Human Bone Marrow-Derived Mesenchymal Stem Cells via ß2-Adrenoceptor-Mediated ERK1/2 and PKA Phosphorylation. Int J Mol Sci 21:N/A (2020). PubMed: 32486305
  • Garza Carbajal A  et al. Depolarization-Dependent C-Raf Signaling Promotes Hyperexcitability and Reduces Opioid Sensitivity of Isolated Nociceptors after Spinal Cord Injury. J Neurosci 40:6522-6535 (2020). PubMed: 32690613
  • Zeng XJ  et al. A2A R inhibition in alleviating spatial recognition memory impairment after TBI is associated with improvement in autophagic flux in RSC. J Cell Mol Med 24:7000-7014 (2020). PubMed: 32394486
View all Publications for this product

Customer reviews and Q&As

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Western blot abreview for Anti-PKA R2/PKR2 (phospho S99) antibody [E151]

 Good
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Jun 23 2009

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