Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E151 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
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 |
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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 |
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ICC/IF |
1/500.
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Notes |
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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. |
Target
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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
modificationsPhosphorylated by the activated catalytic chain. -
Cellular localization
Cytoplasm. Cell membrane. Colocalizes with PJA2 in the cytoplasm and the cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 5576 Human
- Entrez Gene: 19087 Mouse
- Entrez Gene: 29699 Rat
- Omim: 176910 Human
- SwissProt: P13861 Human
- SwissProt: P12367 Mouse
- SwissProt: P12368 Rat
- Unigene: 631923 Human
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
see all
Images
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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) - 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.
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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.
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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 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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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.
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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 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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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
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.
Datasheets and documents
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SDS download
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Datasheet download
References (30)
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