PRODUCT CODE: ET1610-29

Recombinant Phospho PKA R2 (S99) Monoclonal Antibody (ET1610-29)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Pig

ICC staining of Phospho-PKA R2 (S99) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-29, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-PKA R2 (S99) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-29, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-PKA R2 (S99) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-29, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Phospho-PKA R2 (S99) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29, 1/50) for 30 minutes 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 mouse testis tissue using anti-Phospho-PKA R2 (S99) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29, 1/50) for 30 minutes 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 mouse heart tissue using anti-Phospho-PKA R2 (S99) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-29, 1/50) for 30 minutes 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.
ICC staining of Phospho-PKA R2 (S99) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-29, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Pig

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho PKA R2 (S99) Monoclonal Antibody (ET1610-29)

Immunogen

Recombinant protein

Host

Rabbit

Modification

Phospho

Modification Site

S99

Positive Control

Hela, MCF-7, human spleen tissue, mouse testis tissue, mouse heart tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC54-04

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

51 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-PKA R2 (S99)

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; 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

SEQUENCE SIMILARITIES

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

Cell membrane, Cytoplasm.

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.