PKA RIIbeta (AA 1-418) antibody

Details for Product No. ABIN968072
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Antigen
Epitope
AA 1-418
Reactivity
Human
(2), (2)
Host
Mouse
(2)
Clonality (Clone)
Monoclonal ()
Application
Western Blotting (WB), Immunofluorescence (IF)
(2), (2)
Pubmed 6 references available
Quantity 50 µg
Options
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Catalog No. ABIN968072
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Immunogen Human PKA RIIbeta, aa 1-418
Clone DF7
Isotype IgG1
Cross-Reactivity Chicken, Dog (Canine), Mouse (Murine), Rat (Rattus)
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Background CAMP-dependent Protein Kinase (PKA) is composed of two distinct subunits: catalytic (C) and regulatory (R). Four regulatory subunits have been identified: RIalpha, RIß, RIIalpha, and RIIß.These subunits define type I and II cAMP-dependent protein kinases. Following binding of cAMP, the regulatory subunits dissociate from the catalytic subunits, rendering the enzyme active. Type I and type II holoenzymes have three potential C subunits (Calpha, Cß, or Cgamma). Type II PKA can be distinguished by autophosphorylation of the R-subunits, while type I PKA binds Mg/ATP with high affinity. Most cells express both type I and type II PKAs. Although the Ralpha isoforms are ubiquitously expressed, the Rß isoforms are predominant in nervous and adipose tissues. There are indications that the deletion of the gene for PKA RIIß results in lack of long-term potentiation in a select group of hippocampal cells, suggesting an important role for this protein in the neurosciences.
Molecular Weight 53 kDa
Application Notes Methanol Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS.
Triton-X 100 Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS.
Comment

Related Products: ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Supplier Images
anti-PKA RIIbeta (AA 1-418) antibody Western blot analysis of PKA RIIbeta on human endothelial lysate (left). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of PKA RIIbeta.
anti-PKA RIIbeta (AA 1-418) antibody (2) Immunofluorescent staining of SK-N-SH cells (right). Cells were seeded in a 384 well collagen coated Microplates at ~ 8,000 cells per well. After overnight incubation, cells were stained using the Triton X100 fix/perm protocol and the anti- PKARIIb antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen). The image was taken on a Pathway 855 or 435 imager using a 20x objective. This antibody also stained SH-SY5Y and C6 cells using both the Triton X100 and methanol fix/perm protocols.
Product cited in: Skålhegg, Landmark, Foss et al.: "Identification, purification, and characterization of subunits of cAMP-dependent protein kinase in human testis. Reverse mobilities of human RII alpha and RII beta on sodium dodecyl sulfate-polyacrylamide gel electrophoresis compared with rat and bovine R" in: The Journal of biological chemistry, Vol. 267, Issue 8, pp. 5374-9, 1992 (PubMed).

Sandberg, Skålhegg, Jahnsen: "The two mRNA forms for the type I alpha regulatory subunit of cAMP-dependent protein kinase from human testis are due to the use of different polyadenylation site signals." in: Biochemical and biophysical research communications, Vol. 167, Issue 1, pp. 323-30, 1990 (PubMed).

Casey, Vaughan, He et al.: "Mutations in the protein kinase A R1alpha regulatory subunit cause familial cardiac myxomas and Carney complex." in: The Journal of clinical investigation, Vol. 106, Issue 5, pp. R31-8, 2000 (PubMed).

Taskén, Collas, Kemmner et al.: "Phosphodiesterase 4D and protein kinase a type II constitute a signaling unit in the centrosomal area." in: The Journal of biological chemistry, Vol. 276, Issue 25, pp. 21999-2002, 2001 (PubMed).

Tavalin, Colledge, Hell et al.: "Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 22, Issue 8, pp. 3044-51, 2002 (PubMed).

General OSullivan, Kennedy, Casey et al.: "Anabolic-androgenic steroids: medical assessment of present, past and potential users." in: The Medical journal of Australia, Vol. 173, Issue 6, pp. 323-7, 2000 (PubMed).

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