Phospho-PKA R2 (S99) Recombinant Rabbit Monoclonal Antibody [SC54-04]
Catalog# ET1610-29
Phospho-PKA R2 (S99) Recombinant Rabbit Monoclonal Antibody [SC54-04]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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Pig
Predicted species support after-sales service
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unconjugated
Overview
Product Name
Phospho-PKA R2 (S99) Recombinant Rabbit Monoclonal Antibody [SC54-04]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser99 of Human PKA R2 aa 82-126 / 404.
Species Reactivity
Human, Mouse, Rat (Predicted: Pig)
Predicted species support after-sales service
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 46 kDa
Positive Control
PC-12 treated with 100nM Calyculin A for 30 minutes cell lysate, human kidney tissue, mouse kidney tissue, rat kidney tissue, human colon cancer tissue, PC-12 cells treated with 100nM Calyculin A for 30 minutes, PC-12.
Conjugation
unconjugated
Clone Number
SC54-04
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:1,000
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IP
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1-2μg/sample
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FC
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1:1,000
Target
Function
The second messenger cyclic AMP (cAMP) mediates diverse cellular responses to external signals such as proliferation, ion transport, regulation of metabolism and gene transcription by activation of the cAMP-dependent protein kinase (cAPK or PKA). Activation of PKA occurs when cAMP binds to the two regulatory subunits of the tetrameric PKA holoenzyme resulting in release of active catalytic subunits. Three catalytic (C) subunits have been identified, designated Cα, Cβ and Cγ, that each represent specific gene products. Cα and Cβ are closely related (93% amino acid sequence similarity), whereas Cγ displays 83% and 79% similarity to Cα and Cβ, respectively. Activation of transcription upon elevation of cAMP levels results from translocation of PKA to the nucleus where it phosphorylates the transcription factor cAMP response element binding protein (CREB) on serine 133 which in turn leads to TFIIB binding to TATA-box-binding protein TBP1, thus linking phospho-CREB to the pol II transcription initiation complex.
Background References
1. 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).
2. Bantubungi K et al. Cdkn2a/p16Ink4a regulates fasting-induced hepatic gluconeogenesis through the PKA-CREB-PGC1a pathway. Diabetes 63:3199-209 (2014).
Sequence Similarity
Belongs to the cAMP-dependent kinase regulatory chain family.
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.
Post-translational Modification
Phosphorylated by the activated catalytic chain.
Subcellular Location
Cytoplasm, Cell membrane.
Synonyms
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Phospho-PKA R2 (S99) on different lysates with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/2,000 dilution.
Lane 1: PC-12 cell lysate
Lane 2: PC-12 treated with 100nM Calyculin A for 30 minutes cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 46 kDa
Observed band size: 50 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-29) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded human kidney tissue untreated / treated with λpp with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue untreated / treated with λpp with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded rat kidney tissue untreated / treated with λpp with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of PC-12 cells untreated / treated with 100nM Calyculin A for 30 minutes labeling Phospho-PKA R2 (S99) with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-PKA R2 (S99) antibody (ET1610-29) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of PC-12 cells labeling Phospho-PKA R2 (S99).
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-29, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Phospho-PKA R2 (S99) was immunoprecipitated from 0.2 mg PC-12 cell lysate with ET1610-29 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1610-29 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: PC-12 cell lysate (input)
Lane 2: ET1610-29 IP in PC-12 cell lysate
Lane 3: Rabbit IgG instead of ET1610-29 in PC-12 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 20 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
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β2-Adrenergic Receptor Enhances the Alternatively Activated Macrophages and Promotes Biliary Injuries Caused by Helminth Infection
Journal: Frontiers In Immunology
DOI:
IF: 7.562
Application: WB
Reactivity: Mouse
Publish date: 2021 Oct
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